Breast-Cancer reagents: Novus Biologicals

NB100-304

53BP1 Antibody - BSA Free

(14 Reviews)

(817 Publications)

0.1 mg

NB100-304

Irradiated cochlear spiral ganglion cells, mouse, frozen section of fixed material. Image from verified customer review.

<b>Genetic Strategies Validation. </b>53BP1 was detected in immersion fixed HeLa cells (left) but was not detected in 53BP1 knockout Hela cells (right) using Rabbit Anti-human 53BP1 polyclonal antibody (Catalog #NB100-304) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

Total protein from HeLa, A431, Neuro2A, and PC12 was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/mL in 5% block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. The observed molecular weight for these samples are ~250 kDa and the theoretical molecular weight is 214 kDa.

Human breast tumors stained with 53BP1 antibody. Image from verified customer review.

Detection of Human and Mouse 53BP1 by IHC. Sample: FFPE sections of human ovarian carcinoma (left) and mouse teratoma (right). Antibody: 53BP1 Antibody (Catalog #NB100-304) used at a dilution of 1:1000 (1ug/mL). Detection: DAB.

Human medulloblastoma (DAOY) and mouse astrocyte (C8D1A) cell lines were exposed for 48 hours to DMSO or 1ug/mL of the DNA damaging agent EPE. Cells were immunostained for 53BP1 (green). The nuclei were counterstained with DAPI (blue). Image from verified customer review.

An intracellular stain was performed on RH-30 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Immunoprecipitation analysis lysates from HCC44 cells in 1% NP40. Image from verified customer review.

<b>Biological Strategies Validation. </b>53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-304-img0041.jpg

<b>Biological Strategies Validation. </b>53BP1-Antibody-Western-Blot-NB100-304-img0045.jpg

HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with 53BP1 Antibody conjugated to DyLight 550 (NB100-304R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody NB100-304 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Whole cell lysate from U2OS or 293T cells. Bands indicate an observed molecular weight of ~220 kDa and the theoretical molecular weight is 214 kDa.

<b>Biological Strategies Validation. </b>Upper Panel: 53BP1 foci in proliferating MEFs. Lower Panel: 53BP1 foci in proliferating MEFs exposed to 10 Gy of IR.

Embryonic Fibroblast cells pre-extraction for 5 mins with CSK buffer. Fixed with 4% PFA and 75% Ethanol. Primary Antibody at 1:1000. Secondary Antibody at 1:1000. Image from verified customer review.

<b>Biological Strategies Validation. </b>Upper Panel: Control untreated cells. Lower Panel: Cells exposed to irradiation (10 Gy) and probed for 53BP1 foci. Cells were grown on coverslips, fixed with 4% paraformaldehyde, methanol permeabilized, blocked for 1 h, RT. Incubated with primary antibody (1:200) overnight, washed 3x with PBS, probed with tubulin (Alexa Fluor 594) antibody for 2 h, RT. Washed 3x with PBS, mounted on slides using prolong gold, imaged using Nikon confocal microscope (100x oil). Image from verified customer review.

<b>Genetic Strategies Validation. </b>53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-304-img0044.jpg

<b>Biological Strategies Validation. </b>53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-304-img0047.jpg

53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-304-img0046.jpg

Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody NB100-304 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody NB100-304 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Staining of 53BP1 in human colon cancer using DAB with hematoxylin counterstain.

An intracellular stain was performed on A431 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

For 53BP1 staining, samples were incubated for 1 hour with 1:200 53BP1 primary antibody in PBS/BSA 1%, at room temperature. Green: 53BP Blue: DAPI. Image from verified customer review.

<b>Biological Strategies Validation. </b>DYNLL1 is required for 53BP1-dependent p53 responses to Nutlin-3. a Immunoblot analysis of the MCF-7 cell lines used in (a) with anti-53BP1 or anti-RPA32 antibodies prior to N3 treatment. b Quantification of n = 3 independent experiments represented in (a), each performed in triplicate. Mean ± SD. c Indicated parental or stably complemented 53BP1-/- MCF-7 cell lines were incubated in the presence (11 days) or absence (7 days) of Nutlin-3 (4 µM)

<b>Genetic Strategies Validation. </b>Depletion of PNUTS and NPAT induces a global DDR.(A) Induction of telomere dysfunction–induced foci (TIF) upon candidate knockdown in HeLa cells from Fig 5. Depletion of TRF2 was used as a positive control. 53BP1 was detected by indirect IF and telomeres by FISH with a Cy3-[CCCTAA]3 probe. (B) Frequency distribution of the number of TIFs per cell from (A). n indicates the number of cells analyzed for each condition and red lines mark the mean.

<b>Biological Strategies Validation. </b>HPV+ HNSCCs have decreased expression of NHEJ and HR proteins including DNA-Pk and BRCA2Cells were treated with mock or 4 Gy radiation, harvested and lysed at 15 minutes post-treatment, and analyzed by western blot for relative expression of indicated proteins. beta -actin was used as a loading control. Shown is a representative blot from 2 independent experiments.

<b>Biological Strategies Validation. </b>LNCaP cells in culture were treated for 24 hrs with DMSO or ENZ, then irradiated with 0 or 1 Gy.Cell were fixed and immunostained for 53BP1 0.5, 1.5 and 24 hrs after irradiation. Immunofluorescent images were obtained by confocal microscopy. White bar = 20 microns. A) DMSO alone, B) ENZ alone, C) 1 Gy, D) ENZ + 1 Gy. White bar = 20 microns.

Left: Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 1 mg/ml. A specific band was detected for 53BP1 at approximately 353 kDa (as indicated) using both 4 µg/ml and 20 µg/ml of Rabbit Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # NB100-304) followed by HRP-conjugated Goat Anti-Rabbit Secondary Antibody (Catalog # 042-206). This experiment was conducted under reducing conditions and using the 66-440kDa separation system. Right: Simple Western electropherogram showing the same Rabbit Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # NB100-304) tested at 4 µg/ml (blue line) and 20 µg/ml (green line) in the HepG2 human hepatocellular carcinoma cell line.

Representative immunohistochemistry staining for 53BP1 expression in pancreatic adenocarcinoma. (A) is high intensity of 53BP1 expression and (B) is low intensity of 53BP1 expression.

53BP1 was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-53BP1 Antigen Affinity Purified Polyclonal Antibody conjugated to DyLight 650 (Catalog # NB100-304C) (light blue) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Depletion of TPX2 causes defects in 53BP1 ionizing radiation-induced foci formation.(A) The protein level of 53BP1 is not affected by miRNA-mediated depletion of TPX2. Levels of actin used as loading controls. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0110994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Nuclear intensity of key BER proteins.A) Quantification of mean nuclear fluorescence intensity is shown for PARP1 (**** P 25 cells each with Z-stacked (optically sectioned) nuclei.DOI://dx.doi.org/10.7554/eLife.01222.017 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - 53BP1 cofractionates with lamins A/C in HDF. (A) HDF were treated with IR (5 Gy, 1 h) & fractionated by sequential treatments of detergent (CSK/T) & DNase I. Extracts were then processed for immunoblotting using the indicated antibodies. WCE, whole cell extract. P, pellet; S, supernatant. (B) HDF were treated with IR (2 Gy; 1 h) & extracted in situ with CSK/T followed by DNase I digestion before being processed for immunofluorescence using lamin A/C & 53BP1 antibodies. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - 53BP1 focus formation. (A) Bars indicate the mean ± standard error of 53BP1 focus-forming cells in three independent control & three independent Asciz−/− primary MEF cultures under basal conditions (open bars), in response to 24-h aphidicolin treatment (black bars) & following 30 min recovery after ionizing radiation (grey bars). More detailed quantifications of foci/cell are shown in Supp. Fig. S2. (B) Subcelllular distribution of 53BP1 in cell fractionation assays. M601 is used as a mitochondrial marker, & tubulin as a cytosolic marker. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S1568786417301349), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - hsRAD51-II3A expressing cells accumulate 53BP1 foci after treatment with HU. Representative images depicting 53BP1 foci (green) in PCNA (red) positive cells after the indicated treatments. DNA is stained in blue. Scale bar = 10 μm. Dot plot depicts quantitation of 53BP1 foci after indicated treatments. Data is combined data consisting of at least 100 nuclei from two independent experiments. Red line represents the mean. Values are presented as mean ± SD. Statistical significance was determined using the Mann–Whitney U test. Source data are provided as a source data file Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31562309), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Persistent DNA damage exists in ALT cells. (A) IF-FISH showing DNA damage response (53BP1) in ALT (VA13, U2OS, SAOS2, SKLU-1), telomerase positive (MCF7, A549), & human normal fibroblast cells (MRC5, BJ). Scale bar: 10 μm. (B) Quantification of (A). Data represent the mean ± SEM, n = 3. More than 100 cells were counted for each experiment. (C) FACS analysis of apoptosis in cells listed in (A). (D) Quantification of (C). Data represent the mean ± SEM, n = 3. (E) Proliferation of p53-positive (VA13, U2OS) & p53-defictive (SAOS2, SKLU-1) ALT cells. (F) Western blot showing activation of JNK (phosphorylation) & accumulation of c-myc in ALT cells. Human normal BJ cells were used as a control Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31115790), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - LNCaP cells in culture were treated for 24 hrs with DMSO or ENZ, then irradiated with 0 or 1 Gy.Cell were fixed & immunostained for 53BP1 0.5, 1.5 & 24 hrs after irradiation. Immunofluorescent images were obtained by confocal microscopy. White bar = 20 microns. A) DMSO alone, B) ENZ alone, C) 1 Gy, D) ENZ + 1 Gy. White bar = 20 microns. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30933998), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - IMR90 cells depleted for Ajuba undergo a DNA damage response. (A,C,D) Western blots for Ajuba, Cyclin A, Rb, Chk1 phosphorylation, & PARP cleavage on lysates prepared from IMR90 cells 72 h after transfection with siRNA #3. The loading controls with GAPDH for each blot are shown. (B) Staining for p53BP1 in Ajuba-depleted cells (siRNA#3), with siGFP as a control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23755068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - DYNLL1 is required for 53BP1-dependent p53 responses to Nutlin-3. a Immunoblot analysis of the MCF-7 cell lines used in (a) with anti-53BP1 or anti-RPA32 antibodies prior to N3 treatment. b Quantification of n = 3 independent experiments represented in (a), each performed in triplicate. Mean ± SD. c Indicated parental or stably complemented 53BP1-/- MCF-7 cell lines were incubated in the presence (11 days) or absence (7 days) of Nutlin-3 (4 µM) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30559443), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - RNA:DNA hybrids promote 53BP1 foci formation.(a) Immunofluorescence using the anti-flag & anti-53BP1 antibody in cells expressing Flag-tagged hybrid-binding (HB) domain or flag-tagged active RNaseH1. 53BP1 foci were quantified in cells expressing HB-domain or RNaseH1. Column bars represent the mean of n number of cells (described on each column), from three independent experiments. Error bars represent SEM. *P ≤ 0.05; **P 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25582120), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Hypoxia causes DNA damage in patient-derived colonosphere cultures in vitro(a) Two independent patient-derived colonospheres were cultured in normoxia (21%) or hypoxia (0.1%) for 24 hours. Immunofluorescence was then used to asses gamma H2AX (green) in the nuclei DAPI (blue). (b) Bar graphs showing the quantification of gamma H2AX observed in (a) (n=3), shown as percentage of the mean gamma H2AX intensity at normoxia. (c) Representative confocal pictures of the comet assay of colonospheres cultured as in (a). (d) Bar graphs showing the quantification of DNA damage observed with the comet assay (Extend tail moment) (n=73). (e) Colonospheres were cultured as in (a) & gamma H2AX was assessed by FACS. The plots show normoxic gamma H2AX levels in red & hypoxic gamma H2AX levels in blue in two independent cell lines. (f) Human colonospheres were exposed to hypoxia for the indicated periods of time. Cell lysates were then analyzed by Western blotting for the indicated markers (cropped). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.21145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - TRF1HP1 alpha results in reduced TIFs induced by mutant hTR expression. Cells stably expressing TRF1HP1 alpha (WT or mutant variants of HP1 alpha ) were infected with lentivirus containing WT hTR, or mutant hTR (47A or TSQ1) on day 0, selected for stable expression after 48 h, & analyzed on day 5. a Fluorescence microscopy images of representative cells expressing mutant hTR 47A stained with telomeric (Tel-Cy3) peptide nucleic acid (PNA) probes (magenta in merged image) via fluorescent in situ hybridization (FISH), antibody against DNA-damage repair protein marker 53BP1 (green in merged image), & counterstained with DAPI. Zoom-in images (the last row) correspond to yellow-squared regions of the row above. b % TIF per telomere of each nucleus is quantified; n = 94–159 nuclei combining data of three independent experiments. ****p 0.05. (c) Immunofluorescence staining of gamma H2AX-53BP1 positive foci 7 days following exposure to 1Gy of the indicated radiation type. Scale bar is 10 µm. (d) Immunofluorescence staining for BrdU & 53BP1 7 days following exposure to 1Gy Fe ions. Scale bar is 10 µm. The graph represents the average of scoring in 5 different fields a total of 70 cells. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0108234), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - SPOC1 is recruited to I-SceI-induced DSBs where it colocalizes with 53BP1. (A) In situ immunofluorescence studies with U2OS19 ptight13 GFP-LacR cells with a stably integrated I-SceI cleavage site flanked by lac operator repeats reveals localization of the GFP-lac repressor protein (GFP-LacR) at the lac-operator DNA sequences in the nucleus before (−I-SceI) & 16 h after I-SceI-induced (+ I-SceI) DSB. In contrast, SPOC1 (white) & 53BP1 (red) are distributed throughout the nucleus in the absence of I-Scel, & partially colocalized in naturally occurring repair foci. After I-SceI cleavage to generate DSBs, SPOC1 & 53BP1 colocalize at distinct foci, including the cleaved DNA adjacent to DNA-bound GFP-LacR. Proteins were visualized by immunostaining & confocal microscopy. Scale bars = 10 μm. (B) Quantitative analysis of SPOC1 recruitment to the 53BP1 & GFP-LacR positive lacO array before & 16 h after I-SceI induction. (C) Monitoring of the kinetics of SPOC1 & 53BP1 recruitment to DSBs between 0 & 24 h post-I-SceI induction. (D) Quantifying ATM kinase inhibitor-mediated effects on recruitment of SPOC1 & 53BP1 to DSBs as evident 16 h after I-SceI-induced cleavage. (E) SPOC1 & 53BP1 colocalize at a large discrete endogenous repair focus as observed in some non-irradiated U2OS cells by immunostaining. Scale bar = 10µm. Image collected & cropped by CiteAb from the following publication (//academic.oup.com/nar/article-lookup/doi/10.1093/nar/gks868), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - A-type lamin depletion inhibits DNA repair & reduces cellular fitness to DNA damage. (A) Wild-type (WT) or Y259X HDF treated w/ proteasome inhibitor MG-132 (30 μm). Cell extracts prepared & levels of 53BP1 assessed by immunoblotting. beta -tubulin a loading control. WCE, whole cell extract. The relative levels of 53BP1 expressed relative to levels of untreated WT HDF (1.00). In untreated Y259X cells, 53BP1 levels fell to 15% & restored to ∽50% following treatment w/ MG-132. (B) Cell extracts prepared from WT & Y259X HDF & assessed by immunoblotting w/ indicated antibodies. Shown is a representative set of blots from two independent experiments. (C) WT & Y259X HDF biochemically extracted into cytoplasmic & nuclear fractions before immunoblotting for levels of 53BP1. (D) HDF & Y259X cells treated w/ IR (3 Gy), fixed at the times indicated & stained w/ gamma -H2AX antibody. The number of gamma -H2AX foci at each time point quantified & data represents the mean ± SD from 3 independent experiments. (E) U2OS cells transfected w/ siCTRL or siLMNA siRNA & plated onto 60-mm dishes. The cells then exposed to varying doses of IR & fixed 12 days later in a solution containing crystal violet. The surviving fraction at each IR dose calculated based on the plating efficiency of untreated plates. Data represent the mean ± SD from 3 independent experiments. (F) HDF or Y259X cells treated w/ DMSO or KU55933 ATM inhibitor (10 μm) before exposure to indicated doses of IR then fixed, processed & stained w/ gamma -H2AX antibody. Residual gamma -H2AX foci enumerated & the data represent the mean ± SD from 3 independent experiments. (G) HDF or Y259X cells exposed to IR, briefly extracted w/ CSK buffer & processed for IF w/ gamma -H2AX & pKAP-1 antibodies. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - TRF2/Terc double null mutation induces DNA damage response resulting in terminal differentiation of metastatic tumorsHistopathology of primary SCC from K14Cre;TRF2+/+;Terc+/+ A. & K14Cre;TRF2f/f;Terc-/- C. mice is shown by H&E staining. Scale bar = 10 μm. Histopathology of metastatic SCC from K14Cre;TRF2+/+;Terc+/+ B. & K14Cre;TRF2f/f;Terc-/- D., E. mice is shown by H&E staining. Scale bar in (D) = 50 μm. Apoptotic cells in primary SCC from K14Cre;TRF2+/+;Terc+/+ F. & K14Cre;TRF2f/f;Terc-/- G. mice are shown by TUNEL assay. Scale bar = 10 μm. Proliferating cells were detected in K14Cre;TRF2+/+;Terc+/+ H. & K14Cre;TRF2f/f;Terc-/- I. primary SCC by PCNA immunohistochemistry & quantitated in J.. Error bars indicate SEM. Asterisk indicates statistically significant difference. Co-localization of 53BP1 (shown by immunofluorescence, AlexaFluor 488) at telomeres (shown by fluorescence in situ hybridization, Cy3) in histopathologic sections from K14Cre;TRF2+/+;Terc+/+ K. & K14Cre;TRF2f/f;Terc-/- L. primary SCC & quantitated in M. Error bars indicate SEM. Nuclei were counterstained with DAPI. Scale bar = 5 μm. N. Average telomere length ratios in CD34+ stem, Lgr6+ stem, & basal cells from K14Cre;TRF2+/+;Terc+/+ & K14Cre;TRF2f/f;Terc-/- primary SCC were determined by qPCR. Error bars represent SEM. O. Phospho-ATM, phospho-Chk2, & p53 expression in K14Cre;TRF2+/+;Terc+/+ & K14Cre;TRF2f/f;Terc-/- primary SCC is shown by western blot. beta -actin expression was used to control for equal loading of each lane. Representative blots are shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29113290), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Increased Neurogenesis in the Absence of INK4a/ARF Expression Is Independent of Apoptosis & Microglial Activation(A) Quantification of the number of apoptotic cells, as detected by TUNEL, in the DG 6 hr following exposure of WT, ink4a/arf-deficient (INK4a/ARF KO), & p53-deficient (p53 KO) mice to 6 Gy cranial IR. Indicated is the average number of apoptotic cells per DG section (n = 4 mice per group).(B) Quantification of CD68 expression in the DG 24 hr following exposure of mice to IR.(C) Representative confocal images showing CD68+ cells (in red) next to apoptotic TUNEL+ cells (in green) following exposure of WT mice to IR. Note the increase in CD68 signal & side by side localization of CD68+ & TUNEL+ cells following IR.(D–F) Proportion of neurons (NeuN, in green), young neurons (DCX, in green), & progenitor cells (Sox2, in green) showing persistent DNA damage foci (53BP1, in red) in the hippocampus of mice sacrificed 8 weeks post exposure to IR. Nuclei were stained with DAPI.(G) Representative confocal images of brain sections from irradiated WT mice displaying 53BP1 DNA damage foci with the indicated cell markers are shown. Images were acquired by a Leica microsystems TSC SP8 HyVolution confocal microscope & an HC PL APO CS2 40×/1.3 oil objective. The p values (∗p 25 cells each. (E) GamGFP forms IR-induced foci in Ku80-defective MEFs. (F) Zoomed image from E. (G) IR-induced foci containing Gam only, 53BP1 only or both Gam & 53BP1 (>2600 total foci counted in three independent experiments). Error bars, SEM. Scale bars = 5 μm.DOI://dx.doi.org/10.7554/eLife.01222.012GamGFP marks DSBs in mammalian cells.(A) Live analysis of GamGFP localization to laser-induced DNA damage. Hela cells producing GamGFP were laser damaged along the cell track indicated by the red line at 0 min (m) & images were taken at the indicated times as shown. (B) GamGFP co-localizes with gamma H2AX in fixed Hela cells.DOI://dx.doi.org/10.7554/eLife.01222.013Ku inhibits GamGFP recruitment at DSBs independently of non-homologous end joining.Cells lacking either Ku80 or LigIV are defective in non-homologous end joining (NHEJ), yet the presence of Ku still inhibits recruitment of GamGFP to laser-induced DSBs even in NHEJ-defective cells, & thus independently of the cell's ability to complete NHEJ. Whereas it could have been possible that reduced GamGFP recruitment in the presence of Ku was caused by reduced persistence of DSBs due to their repair by NHEJ, our data show instead that Ku inhibits recruitment independently of successful NHEJ & imply that Ku binding to DSEs itself is inhibitory.DOI://dx.doi.org/10.7554/eLife.01222.014 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Loss of functional PRPF31 induce RNA:DNA hybrid dependent genomic instability but not in mice retinal neurons.(a andb) gamma H2AX & 53BP1 foci analysis in PRPF31 siRNA-transfected RPE-1 cells. (c & d) gamma H2AX & 53BP1 foci analysis in vasculo-stromal fraction derived primary cells fromPrpf31+/A216P mice (Prpf31-ki). (e) gamma H2AX & 53BP1 foci analysis in retina fromPrpf31+/A216P mice on postnatal day 20. All column bars represent the mean. For (a-d) “n”, mentioned on respective column, signify number of cells analyzed from two independent experiments. For (e) n=16 for each column & signify number of retinal sections analyzed; acquired from n=4 eyes. Error bars represent Standard error of Mean (SEM). *P≤0.05; **P 0.9999, ****P 0.05). ns = not significant. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.21145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. (A) Left; HDF were subjected to IR (10 Gy) & allowed to recover for 1 h. Association between A-type lamins & 53BP1 was assessed by immunoprecipitation of endogenous lamin A/C followed by immunoblotting with 53BP1 & lamin A/C antibodies. WCE, whole cell extract, IP: immunoprecipitates. WCE represents 5% input. Right; endogenous lamin A/C was immunoprecipitated & supernatants or pellets were analysed by immunoblotting for interaction with JMJD2A. (B) U2OS/GFP-lamin A cells were pretreated with caffeine (20 mm) for 1 h before exposure to IR (10 Gy, 1 h recovery). Cell extracts were then subjected to immunoprecipitation using GFP-Trap beads, & bound complexes were then analysed by immunoblotting using 53BP1 & GFP antibodies. WCE represents 1% input. (C) As in (C) except cells were pretreated with 10 μm ATMi for 1 h before IR. WCE represents 1% input. (D) U2OS/GFP-lamin A cells were subjected to laser micro-irradiation, fixed 1 h later & immunostained with gamma -H2AX antibody. Scale bar, 10 μm. (E) U2OS cells were transfected with siCTRL or siLMNA & subjected to laser micro-irradiation, fixed 15 min later & then processed for immunofluorescence with gamma -H2AX & 53BP1 antibodies. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Persistence of repair foci at suboptimal pHe. (A) Left panel shows immunofluorescence detection of gamma H2AFX & 53BP1 following 24 h recovery from bleomycin treatment. Control, sample before damage induction. Colocalization is shown in white; PCC, Pearson correlation coefficient score. Right panel corresponding to gamma H2AFX foci/cell at 24 h & 48 h following recovery. Only foci larger than 10 pixels were counted. Experiments were performed in triplicate. Values are mean ± SEM. *P 1.0 mm, >2.0 mm) are shown. Data are expressed as the mean ± SD (n = 3, *p 10 foci was called positive & counted). DAPI was used for nuclear staining; (B) The relative DNA damage in primary-culture tumor cells at 120 min post-IR was analyzed by comet assay (upper), & gamma H2AX foci formation in primary-culture tumor cells at 24 h post-IR was presented，“+” & “−“ indicated whether VPA was added in the groups (lower left) & calculated (lower right); (C) The clonogenic survival assay was used to detect survival in the U2OS cells treated with the combination of 0.5 or 1.0 mM VPA with 10 μM ABT888 (PARPi). Each data point in the graphs was from three independent experiments (mean ± SD). p-Values were calculated by Student’s t-test (* p 25 cells each. (E) GamGFP forms IR-induced foci in Ku80-defective MEFs. (F) Zoomed image from E. (G) IR-induced foci containing Gam only, 53BP1 only or both Gam & 53BP1 (>2600 total foci counted in three independent experiments). Error bars, SEM. Scale bars = 5 μm.DOI://dx.doi.org/10.7554/eLife.01222.012GamGFP marks DSBs in mammalian cells.(A) Live analysis of GamGFP localization to laser-induced DNA damage. Hela cells producing GamGFP were laser damaged along the cell track indicated by the red line at 0 min (m) & images were taken at the indicated times as shown. (B) GamGFP co-localizes with gamma H2AX in fixed Hela cells.DOI://dx.doi.org/10.7554/eLife.01222.013Ku inhibits GamGFP recruitment at DSBs independently of non-homologous end joining.Cells lacking either Ku80 or LigIV are defective in non-homologous end joining (NHEJ), yet the presence of Ku still inhibits recruitment of GamGFP to laser-induced DSBs even in NHEJ-defective cells, & thus independently of the cell's ability to complete NHEJ. Whereas it could have been possible that reduced GamGFP recruitment in the presence of Ku was caused by reduced persistence of DSBs due to their repair by NHEJ, our data show instead that Ku inhibits recruitment independently of successful NHEJ & imply that Ku binding to DSEs itself is inhibitory.DOI://dx.doi.org/10.7554/eLife.01222.014 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - DNA repair response to irradiation-induced DNA-breaks in adult mice retina.(a) Immunofluorescence performed using anti-gamma H2AX & 53BP1 antibodies in mice retina 1hr after exposure to 5 Gyrase of ionizing radiation. gamma H2AX appears in all cell types in response to DNA breaks. 53BP only observed in the ganglion cell layer (GC) & inner strata of inner nuclear layer (INL) (seeFigure 1a). Zoom in show foci formation in cells expressing 53BP1. (b) H2AX phosphorylation in all retinal cell types is euchromatin specific. (c) Kinetics of DNA repair in mice retina. Post-Irradiation, mice were sacrificed at indicated times & retinal sections were analysed for gamma H2AX & 53BP1. Left panel show all nuclear layers of retina;right panel show zoom in images to emphasize foci formation. Two mice were used for each condition, which were processed & stained together. Random image were taken using confocal microscope on each eye. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30345028), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Detection of 53BP1 in CTCs from MBC patients in the DETECT IV trial. (a) Selection of MBC patients with CTC-positivity (HER2-) & primary TNBC or primary HER2-, HR+ breast cancer for Eribulin monotherapy & blood sample collection. Blood samples were drawn before therapy (baseline visit), four, twelve, & 24 weeks after the start of Eribulin treatment (1st, 2nd, & 3rd treatment visit) as well as one year after treatment initiation or during premature termination due to disease progression (final visit). The numbers of patients recruited at the different visits are indicated by black columns, the numbers of CTC+ & CTC- patients among them by red & grey columns, respectively. Note that not all of the patients described in Table 1 entered the study during the baseline visit but during later visits which explains the differences in patient numbers. (b) Evaluation of 53BP1 signals in CTCs. Following EpCAM-based CTC enrichment & immunofluorescence microscopic imaging (Cellsearch®), CTCs (CK+, CD45-) were enumerated & 53BP1-staining intensity assessed per individual CTC as indicated by representative examples. A 53BP1 score per patient sample was calculated from the percentage of CTCs without & with light, moderate or strong staining. PBMC, peripheral blood mononuclear cell (CK-, CD45+). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32283863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Treatment w/ exogenous POT1 protein protects LT-HSCs in culture. a Effect of MTM-POT1a on colony formation of HSCs. LT-HSCs (8 week-old) cultured w/ MTM-POT1a for 2 weeks. After culture, Lin− cells isolated & re-cultured in MethoCult™ GF M3434 medium (200 cells per dish). Data are expressed as the mean ± SD (n = 3, *p < 0.01 by Tukey’s test). Representative data from two independent experiments are shown. b, c LT-HSCs (8 week-old) cultured w/ MTM-POT1a or control MTM protein. b Number of total cells & LT-HSCs on day 4, 7, & 10 of culture. Data are expressed as the mean ± SD (n = 6, *p < 0.01 by t-test). c Percentage of Annexin V+PI+ apoptotic cells in LT-HSCs on day 4, 7, & 10 of culture. Data are expressed as the mean ± SD (n = 6, *p < 0.01 by t-test). d LT-HSCs (8 week-old) transduced w/ control shRNA or shPot1a-1. After 2 days of shRNA transduction, GFP+LT-HSCs re-sorted & cultured w/ MTM-POT1a or control MTM protein. After 1 week of culture, Annexin V assay performed. Percentage of Annexin V+PI+ apoptotic cells in GFP+LT-HSC fraction is shown. (n = 3, **p < 0.05 by Tukey’s test). e Immunocytochemical staining of TRF (green), 53BP1 (red), & TOTO3 (blue) in 8 week-old LT-HSCs cultured for 3 weeks (left). Frequencies of TIFs after 3 weeks of culture (right). Data are expressed as the mean ± SD (n = 80–100, *p < 0.01 by t-test). f Representative FACS profiles of 8 week-old LT-HSCs after 3 weeks of culture w/ control MTM protein or MTM-POT1a (left). Numbers of LSK cells & LT-HSCs after 3 week culture (starting from 2400 cells) are shown in right panels. Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative data from five independent experiments are shown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28986560), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Induction of senescence in hepatocytes causes mitochondrial dysfunction & reduces fatty acid oxidation capacity.Hepatocytes isolated from wild-type C57Bl/6 mice acquire a senescent phenotype 6 days after 10 Gy X-ray irradiation: (a) representative images showing SA-beta -Gal in non-irradiated cells (NOIR) & 6 days after irradiation, Merge: light blue=DAPI, dark blue=SA-beta -Gal, scale bar, 20 μm; (b) frequencies of SA-beta -Gal-positive hepatocytes & (c) representative images showing 53BP1 staining in NOIR & 6 days after irradiation. Merge: blue=DAPI, red=53BP1, scale bar 10 μm; (d) average number of 53BP1 foci per cell are significantly increased 6 days after irradiation. (e) Representative images showing Nile red staining in NOIR & 6 days after IR. Merge: blue=DAPI, red=Nile red, scale bar scale bar 20 μm. (f) Average fluorescence intensity of Nile red staining in senescent or non-senescent hepatocytes. (g) Change in OCR in senescent or non-senescent hepatocytes after addition of the fatty acid palmitate as substrate; (h) change in OCR in senescent or non-senescent hepatocytes after inhibition of fatty acid oxidation by etomoxir. Data in b,g,h are mean±s.e.m. & in d,f are mean±s.d. with 3–4 animals per group. Significant differences (t-test) are indicated with *P≤0.05 & **P≤0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28608850), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

An intracellular stain was performed on U-2 OS human osteosarcoma cell line with Rabbit anti-53BP1 Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor ® 647 (Catalog # NB100-304AF647, blue histogram) or matched control antibody (Catalog # NBP2-24981AF647, orange histogram) at 2.5 µg/mL for 30 minutes at RT.

53BP1 was detected in immersion fixed SJCRH30 human Rhabdomyosarcoma cell line using Rabbit anti-53BP1 Antigen Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB100-304AF488) (green) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Reactivity:

Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Pm, Po, Pm, Fe, Pm-Or, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Pm, Po, Pm, Fe, Pm-Or, Rb, Sh

Applications:

, , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-304AF350
Alexa Fluor 405	NB100-304AF405
Alexa Fluor 488	NB100-304AF488
Alexa Fluor 532	NB100-304AF532
Alexa Fluor 594	NB100-304AF594
Alexa Fluor 647	NB100-304AF647
Alexa Fluor 700	NB100-304AF700
Alexa Fluor 750	NB100-304AF750
Alexa Fluor Plus 405	NB100-304AFP405
Alexa Fluor Plus 488	NB100-304AFP488
Alexa Fluor Plus 555	NB100-304AFP555
Alexa Fluor Plus 594	NB100-304AFP594
Alexa Fluor Plus 647	NB100-304AFP647
Alexa Fluor Plus 680	NB100-304AFP680
Allophycocyanin	NB100-304APC
Allophycocyanin/Cy7	NB100-304APCCY7
Biotin	NB100-304B
CoraFluor 1	NB100-304CL1
DyLight 350	NB100-304UV
DyLight 405	NB100-304V
DyLight 488	NB100-304G
DyLight 550	NB100-304R
DyLight 594	NB100-304DL594
DyLight 650	NB100-304C
DyLight 680	NB100-304FR
DyLight 755	NB100-304IR
FITC	NB100-304F
HRP	NB100-304H
Janelia Fluor 525	NB100-304JF525
Janelia Fluor 549	NB100-304JF549
Janelia Fluor 646	NB100-304JF646
Janelia Fluor 669	NB100-304JF669
PE	NB100-304PE
PE/Atto594	NB100-304PEATT594
PE/Cy5.5	NB100-304PECY55
PE/Cy7	NB100-304PECY7
PerCP	NB100-304PCP
mFluor Violet 450 SE	NB100-304MFV450
mFluor Violet 500 SE	NB100-304MFV500
mFluor Violet 610 SE	NB100-304MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-304SS

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NB100-142

Mre11 Antibody - BSA Free

(4 Reviews)

(252 Publications)

0.05 ml

NB100-142

<b>Genetic Strategies Validation. </b>Western Blot: Mre11 Antibody [NB100-142] - Mre11 and NBS1 contribute to OriP replication and EBV episome stability. D98/HR1 cells were transfected with siRNA for Mre11 or NBS1, or Luciferase control and assayed by immunoblot (IB) with antibodies specific for Mre11 (left pane) or NBS1 (right panel), or PCNA (lower panel) as a loading control. Image collected and cropped by CiteAb from the following publication (//journals.plos.org/plosone/article?id=10.1371/journal.pone.0001257) licensed under a CC-BY license.

Immunohistochemistry: Mre11 Antibody [NB100-142] - Immunohistochemical staining of MRE11 in a human epidermis cross section.

Western Blot: Mre11 Antibody [NB100-142] - Western blot analysis of Mre11 on 50 ug of HeLa and MEF lysates, displaying bands at the molecular weight of 81 kDa.

Immunocytochemistry/Immunofluorescence: Mre11 Antibody [NB100-142] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Mre11 [NB100-142] at a 1:500 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Chromatin Immunoprecipitation: Mre11 Antibody [NB100-142] - Aggregated END-seq signal and MRE11 ChIP-seq RPM in WT (left) and Atm -/- (right). To fairly compare ChIP-seq signal between WT and Atm -/-, MRE11 scale is proportional to spike-in normalized END-seq RPM for each genotype. Individual hotspot examples (chr12:34,592,264-34,598,265) are shown below. Note that decreased MRE11 coverage is observed within NDR of Atm -/-. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32051414/) licensed under a CC-BY license.

<b>Genetic Strategies Validation. </b>Western Blot: Mre11 Antibody [NB100-142] - TIG-1 human primary fibroblasts, whole cell lysate (30 ug). Image from verified customer review.

Western Blot: Mre11 Antibody [NB100-142] - MRE11 associates with the DS region of OriP.A) DNA affinity purification assays were performed with Raji nuclear extracts & DNA templates derived from DS wt, DS nm-, or delta DS. Input & template bound proteins were analyzed by Western blot with antibodies to TRF2, MRE11, EBNA1, PCNA, & PARP1, as indicated. B) Schematic of template DNA used for DNA affinity purification in A, & their respective protein binding sites. C) ChIP assays to monitor EBNA1, TRF2, & MRE11 binding at OriP wt or OriP nm- in transfected 293 cells. Immunoprecipitated DNA was quantified by real time PCR relative to total input DNA recovered for each transfected cell. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18040525), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Mre11 Antibody [NB100-142] - Immunoblotting analysis of IR-induced AKT (Ser473) activation. (A) The levels of AKT activation were measured by AKT Ser473 phosphorylation in 293T, 293T/eIF3f & 293T/eIF3f-hMSH4 cells treated with 10 Gy IR in comparison to untreated controls. Levels of p53 Ser15 phosphorylation & the total protein levels of AKT, hMRE11 & p53 were also analyzed. alpha -tubulin was used as a loading control. (B) The levels of Chk2 activation (Chk2 Thr68 phosphorylation) in 293T, 293T/eIF3f & 293T/eIF3f-hMSH4 cells in response to 10 Gy IR. Cell lysates were prepared at 1 hr post IR treatment. Untreated cells were analyzed as controls, while alpha -tubulin was used as a loading control. Image collected & cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-12-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Mre11 Antibody [NB100-142] - The DNA damage response at telomeres uncapped through TRF2 depletion does not require BRCA1 or CtIPImmortalized Brca1F/− MEFs were infected with retroviruses expressing the indicated shRNAs and/or Cre recombinase, followed by selection with puromycin for 72 h. Cell extracts were prepared 48 h later & analysed by Western blotting as indicated. SMC1 & tubulin were used as loading controls. *non-specific band.Cells treated as in (A) were fixed 48 h after selection & stained with an anti-53BP1 antibody (green). Telomeres were visualized with a Cy3-conjugated (CCCTAA)3-PNA probe (red). Yellow arrowheads point to 53BP1 foci that co-localize with telomeres.Quantification of TIFs in cells treated as in (B). A minimum of 200 nuclei were scored for each sample. Error bars represent SD of two independent experiments. P-values were calculated using an unpaired two-tailed t-test. *P ≤ 0.05; NS, P > 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25582120), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Mre11 Antibody [NB100-142] - MRE11 & NBS1 contribute to OriP replication & EBV episome stability.A) D98/HR1 cells were transfected with siRNA for MRE11 or NBS1, or Luciferase control & assayed by immuno-blot (IB) with antibodies specific for MRE11 (left pane) or NBS1 (right panel), or PCNA (lower panel) as a loading control. B) Transient DNA replication assays of OriP-containing plasmids were analyzed by Southern blot. D98/HR1 cells were transfected with OriP plasmid & siRNA for either MRE11, NBS1, or luciferase control (as indicated above each lane). Hirt extracted plasmid DNA was digested with DpnI plus BamHI (top panel) or BamHI only (lower panel) & probed with OriP specific probes. DNA was quantified by PhosphorImager analysis & the replication value was determined as the ratio of DpnI/BamHI to BamHI recovered products. Quantification shown below is a summary of at least four independent replication assays (data not shown). C)) EBV episomes from latently infected D98/HR1 cells were analyzed by pulse field electrophoresis & Southern blotting after transfection of siRNA for control (siLuc), MRE11, or NBS1. Raji cells were used as a control for EBV episome size & abundance. The percentage of EBV episomes relative to the EBV DNA retained in the well is calculated below. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18040525), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Mre11 Antibody [NB100-142] - Evidence that MRE11 & NBS1 are required for EBV episome maintenance.A) OriP plasmid maintenance assays were performed in MRE11 mutant (ATLD3) & reconstituted (ATLD3/wtMRE11) cell lines (left panel) or NBS1 mutant (GM7166) & reconstituted (GM7166/wtNBS1) cell lines (right panel). Plasmids containing OriP & EBNA1 were monitored by Southern blotting of Hirt lysates at 1 (lower panel) & 7 (upper panel) days post-transfection. Phosphorimager quantification of four independent experiments as shown in panal A where maintenance is measured as the ratio of day 7 to day 1 for OriP plasmid detection is shown below. B) EBV transformed B-lymphocytes were analyzed by PFG & Southern blotting for the presence of episomal forms of the EBV genome. Raji, Namalwa, NBS1 (GM15808), NBS1 (GM07078), LCL3472, & LCLAW7 DNA was loaded at equal concentrations & analyzed by ethidium staining of PFG (lower panel) or by Southern blot (upper panel) with EBV specific probe. C) Western blot of the same cell lines used for PFG analysis shown in panel B was probed with antibodies specific for NBS1 (antibody 143 & antibody 277), MRE11, or loading control Actin. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18040525), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Mre11 Antibody [NB100-142] - Involvement of CBP & p300 in the transcription of the BRCA1 & RAD51 genes.(A) Down-regulation of BRCA1 & RAD51 proteins upon depletion of CBP and/or p300. H1299 cells were transfected for 48 hr with non-targeting (siNT), CBP (siCBP), p300 (sip300), or CBP+p300 (siC+p) siRNAs. The cells were harvested & whole cell extracts were subjected to immunoblotting. (B, C) Reduction of BRCA1 & RAD51 transcripts in CBP- & p300-depleted cells. H1299 cells were transfected for 48 hr with non-targeting (siNT), CBP (siCBP), p300 (sip300), or CBP+p300 (siC+p) siRNAs. Cells were harvested & subjected to quantitative real-time PCR for the detection of BRCA1 (B) & RAD51 (C) mRNAs. Expression levels were normalized against the levels of GAPDH mRNA. Data represent the mean ± SD. (D) H1299 cells were transfected for 48 hr with non-targeting (siNT), CBP (siCBP), p300 (sip300), or CBP+p300 (siC+p) siRNAs, & stained with propidium iodide (PI). The percentage of cells in each cell cycle phase was determined by FACS. Percentages of cells in G1, S, & G2/M are shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23285190), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Normal brain and the cerebellum of Mre11-PC delta mice. (A) Abdominal view of wild type (Ctr) and Mre11-PC delta mice showed normal limbs stretching. (B) Bodyweight of control (Ctr) and Mre11-PC delta mice at 18 months of age. The number of mice analyzed is indicated within the graph bars. Error bars indicate the value mean +/- SEM; n.s., not significant; data comparison was performed through Student’s t-test. (C) A dorsal view of the whole brain of control (Ctr) and Mre11-PC delta mice. (D) Brain weight of 12-month-old Ctr and Mre11-PC delta mice. The number of mice analyzed is indicated within the graph bars. Error bars indicate the value mean +/- SEM; data comparison was performed by Student’s t-test. n.s., not significant. (E) Sagittal cryosections of 18-month-old Ctr and Mre11-PC delta mice after staining with anti-MRE11 antibody and DAPI. Note a reduction of MRE11 signal in mutant Purkinje cells. The Purkinje cell layer in the cerebellum region is shown. Dotted lines highlight the nucleus of Purkinje cells. (F,G)Mre11-PC delta mice have normal density and morphology of Purkinje cells. Sagittal sections of control (Ctr) and Mre11-PC delta mice were stained with Calbindin-D28K (D28K) antibody (green) and DAPI (blue) to label Purkinje cells and nuclei, respectively (F). The upper panels show the whole cerebellum, and the lower panels show the magnified view of red frames in the respective upper panels. ML, molecular layer; PCL, Purkinje cell layer. (G) Quantification of total Purkinje cell number in each individual cerebellar lobule from panel (F). Three mice of each genotype were analyzed. Error bars indicate the value mean +/- SEM; data comparison was performed through Student’s t-test. n.s., not significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35153719), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Deletion of Nbs1 is compatible with mouse survival and cerebellar development. (A) Bodyweight of 20∼24 months old mice from the indicated group. (B) Brain weight of the animal for panel (A), Error bars indicate the value mean +/- SEM; n.s., not significant. Data comparison was performed through Student’s t-test. (C,D) Deletion of Nbs1 in Purkinje cells leads to dislocation of MRE11 and RAD50. Sagittal cryo-sections of control (Ctr) and Nbs1-PC delta mice at the indicated age were stained with anti-MRE11 (C) and anti-RAD50 (D) antibodies together with DAPI to label nuclei. The Purkinje cell layer in the cerebellum region is shown. Scale bars: 20 μm. Representative images of sections from two mice are shown. Arrows point to Purkinje cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35153719), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ch, Ha, Bt, Bv, Ca, Pm, Eq, Fe, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Ch, Ha, Bt, Bv, Ca, Pm, Eq, Fe, Pm

Applications:

, , , , , , , , , , ,

Applications: , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

Available Formulations

Formulation	Catalog #	Availability	Price
Purified	NBP3-25349-0.025mg
Purified	NBP3-25349-0.05mg
BSA Free	NB100-142SS

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NB100-182

FANCD2 Antibody - BSA Free

(8 Reviews)

(246 Publications)

0.05 ml

NB100-182

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Resistance of FANCA-mutant cells with defective FANCD2 function to hydrogen peroxide (H2O2). Representative images illustrating staining for subnuclear FANCD2 foci in isogenic fibroblast pairs, either deficient for FANCD2 (PD20) or FANCA (PD220) and their respective wild-type (wt) complemented counterparts. Foci were visualized three hours after treatment with H2O2 (25 uM for 2 hours). Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2008/821529/) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AMPK-activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2-deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with FANCD2 Antibody, phospho-AMPKalpha1 (T172), and AMPKalpha and beta-actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub-FANCD2) is indicated by an arrow. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.

Simple Western: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Simple Western lane view shows a specific band for FANCD2 in 0.1 mg/ml of HeLa lysate using FANCD2 Antibody. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Analysis using the Biotin conjugate of FANCD2 Antibody. FANCD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. Proliferating SiHa cells were exposed to 10 Gy of IR and double color immunofluorescence staining was performed after 8 h. Images were captured in a Kodak digital image system on a Leica fluorescence microscope.

<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AICAR-induced FANCD2 monoubiquitination is dependent on AMPK. Inhibitor of AMPK blocks AICAR-induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 um of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 Antibody to visualize monoubiquitinated FANCD2 (Ub-ANCD2). Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The FA-BRCA Pathway Is Activated on the Sex Chromosomes during Meiosis. Schematic of the FA-BRCA pathway. FA proteins analyzed in this study are shown in color. Western blot analysis with three independent anti-FANCD2 antibodies (G33, E33, and Novus NB100-182 antibody: NB). K561R, PD20 cells expressing a mutated form of FANCD2 incapable of monoubiquitination; WT, PD20 cells complemented with wild-type FANCD2; Vector, PD20 cells containing empty vector. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2211124716313298), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - HES1K109E mutation does not impair MMC-induced HES1 foci formation. HeLa cells transfected with HA-tagged HES1 or HES1K109E coding vectors were processed for immunofluorescence 16 h following treatment with MMC (120 ng/ml). Cells were double-stained with anti-HA and anti-FANCD2 antibodies (mouse anti-HA at 1:500; rabbit anti-FANCD2, Novus Biologicals NB100-182 at 1:2000) followed with secondary antibodies (Goat anti-mouse Alexafluor-555, ThermoScientific A-32727 and goat anti-rabbit Alexafluor-488, ThermoScientific, A-11008). Cell nuclei were labelled with TO-PRO-3 Iodide stain (1:400, ThermoScientific, T3605). Cells were visualized at 100x magnification. Image collected and cropped by CiteAb from the following publication (//bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3243-7), licensed under a CC-BY license.

Immunohistochemistry: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The tissue sections were incubated with a primary antibody cocktail of FANCD2 Antibody (NB100-182) at a dilution of 1:1000 & a monoclonal anti-Ki67 mouse antibody at a dilution of 1:150 for 1 h at room temperature. Then the slides were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG and Alexafluor 594 donkey anti-mouse secondary. The sections were mounted on glass slides using a DAPI-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA), then analyzed under a fluorescence microscope. FANCD2 (top) foci positive NSCL tumor, and FANCD2 (bottom)foci negative NSCL tumor. Magnification: 1000x. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license.

Western Blot: FANCD2 Antibody [NB100-182] - Analysis of FANCD2 (Molecular weight: 164.1 KDa) using the HRP conjugate of FANCD2 Antibody (lot C) in HeLa WCE.

Western Blot: FANCD2 Antibody [NB100-182] - Functional assessment of Fanconi anemia pathway. Western blot with FANCD2 Antibody of non-FA control lymphoblasts (LCL), proband LCL-A-2017, proband LCL-B-2017, FANCB mutant (null) LCL, and FANCD2 mutant (null) LCL. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/mgg3.350), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: FANCD2 Antibody [NB100-182] - Mouse Embryonic Fibroblasts Untreated and treated with Mitomycin C for 24hr with FANCD2 Antibody diluted 1:500 in 15%BCS+0.3% Triton X 100 in PBS. This image was submitted via customer Review.

Immunohistochemistry: FANCD2 Antibody [NB100-182] - Staining of human prostate, glandular epithelium using FANCD2 Antibody. Image using the Biotin format of this antibody.

Representative images of the cellular response to RS (induced by a low dose of hydroxyurea; HU) in U2OS cells, as determined by IF staining for gamma H2AX and FANCD2A selected cell is defined by the yellow arrow, and is enlarged in the bottom panel. Scale bars are indicated.

FANCD2 was detected in immersion fixed A431 human skin carcinoma cell line using Rabbit anti-FANCD2 Antigen Affinity Purified Polyclonal Antibody conjugated to DyLight 550 (Catalog # NB100-182R) (red) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Western blot analysis of FANCD2 & ERCC1 expression in lung cancer cell H1299 following treatment with veliparib. PAR protein level was reduced at 6 h post veliparib 5 μM exposure & maintained at low levels through 48 h in the H1299 cells. However, FANCD2 & ERCC1 protein expression raised simultaneously in these cells post treatment with veliparib. The data suggest that veliparib induces compensatory activation of alternative DNA-repair pathways. C is control (without treatment). Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Workflow for MS-based quantification of CFS associated proteins. (A) Experimental workflow for SILAC-based quantitative MS identification of CFS associated proteins. (B) Flow cytometry analysis of cell cycle distribution of SILAC labeled cells used for enrichment of CFSs as illustrated in (A). (C) IF of FANCD2 & EdU incorporation to assess formation of FANCD2 foci at late replicating regions with & without APH treatments. Cells were synchronized as in (A) & (B). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31180492), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The response to replication stress is a defining feature for Ewing sarcoma compared to the PHATE_1-high developmental context. (A) Western blot showing expression of Fanconi Anemia proteins FANCA, FANCD2, & FANCI in TC32 (Ewing sarcoma cell line) compared to IMR90 (fibroblast cell line); (B) Bar-plot showing the impact of FANC genes knockdown on cell viability in TC32 compared to IMR90; (C) Western blot showing the expression of FEN1 in Ewing sarcoma cell lines TC32, EWS502, & CHLA10 compared to IMR90; (D) Line plot showing the viability of Ewing sarcoma cells compared to IMR90 with increasing doses of FEN1 inhibitor. (** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32290418), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Activation of FANCD2 monoubiquitination following treatment with the EZH2 inhibitor EPZ-6438(A-C) MCF10A (A), HCT116 p53+/+ & p53-/- (B), & U2OS (C) cells were treated with the indicated concentrations of the EZH2-specific inhibitor EPZ-6438 for 24 h (A & B) or 24, 48, & 72 h (C). Whole-cell lysates were prepared & immunoblotted with the indicated antibodies. L.E., light exposure; D.E., dark exposure; L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCI; or FANCD2 RBI, relative band intensity. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Inhibition of class I & II HDACs attenuates FANCD2 & FANCI monoubiquitination in BJ-TERT cells(A) & (B), HeLa cells were pre-treated with the indicated concentrations of trichostatin A (TSA) (A) or vorinostat (SAHA) (B) for 4 h, followed by co-incubation with (+) & without (-) 200 nM MMC for a further 20 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCI, anti-CHK1 pS345, anti-CHK2 pT68, anti-H4K16ac, & anti-alpha -Tubulin antibodies. (C) & (D), BJ-TERT cells were treated identically to that described for HeLa cells above. L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCD2 or FANCI. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Representative images of the cellular response to RS (induced by a low dose of hydroxyurea; HU) in U2OS cells, as determined by IF staining for gamma H2AX & FANCD2A selected cell is defined by the yellow arrow, & is enlarged in the bottom panel. Scale bars are indicated. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.16940), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Functional evaluation of FANCBWT & mutant cDNA. Proband fibroblasts, FANCB mutant (null) fibroblasts & BJ control fibroblasts were HPV16 E6E7 transformed. Either empty vector, wild‐type FANCBcDNA, or mutant FANCBcDNA (p.A319*) was introduced into proband fibroblasts & FANCB mutant (null) fibroblasts. After puromycin selection, cells were cultured with or without MMC 1 μM for 24 hr, after which cells were harvested for the FANCD2 & HA western blot assays Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - BRD4770-induced activation of the FA pathway may occur via inhibition of the PRC2 complex(A) U2OS cells were incubated in the absence (NT) or presence of BRD4770, DZNep, UNC0646, & DZNep & UNC0646 combined (4 μM each) for 24 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCI, anti-EZH2, & anti-alpha -Tubulin antibodies. (B) U2OS cells were incubated in the absence or presence of 2, 5, & 10 μM BRD4770 for 24, 48, or 72 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCI, anti-CHK1 pS345, anti-EZH2, anti-H3K27me3, & anti-alpha -Tubulin antibodies. L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCI; or FANCD2 RBI, relative band intensity. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Chemical library screening for small molecules that inhibit the Fanconi anemia pathway. (A) Schematic of the screening for small molecules that inhibit IR-induced FANCD2 foci formation. (B) Representative photomicrographs of EGFP-FANCD2 foci in PD20F-EGFP-FANCD2 cells untreated & treated with the indicated compounds at the indicated concentration. The cells were treated with compounds immediately before irradiation (15 Gy), & fixed after 12 hours. (C) FANCD2 foci in HeLa cells untreated & treated with the indicated compounds at the indicated concentration. The cells were fixed 8 hours after irradiation (10 Gy) & immunostained with anti-FANCD2 antibody. Scale bar = 20 μm. Image collected & cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-26), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AMPK‐activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. (A) AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2‐deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with anti‐FANCD2, phospho‐AMPK alpha 1 (T172), & AMPK alpha & beta ‐actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub‐FANCD2) is indicated by an arrow. (B) AICAR treatment induces formation of FANCD2 nuclear foci in GM00637I fibroblasts. Cells grown on coverslips in 12‐well plates were treated with 0.25 mm AICAR for 24 h. Cells were immunostained with FANCD2 antibody & Alexa 488‐conjugated anti‐rabbit secondary antibody. FANCD2 foci were visualized by confocal microscopy. Representative images are shown at the top. The number of foci per cell was counted & plotted for ≥ 20 cells (bottom panel). The values represent the mean ± SEM (Student's t‐test, ***P 5 gamma H2AX foci were scored. At least 300 nuclei were scored for each treatment & this experiment was performed three times with similar results. Error bars represent the standard errors of the means from three independent experiments. (B) BJ-TERT cells were incubated with (+) & without (-) 0.4 μM etoposide (VP-16), in the absence or presence of 10 μM BRD4770, 5 μM GSK-J1 & 2.5 μM BIX01294, for 24 h. Whole-cell lysates were immunoblotted with anti-gamma H2AX & anti-PCNA (loading control) antibodies. (C) U2OS cells were incubated in the absence (NT) or presence of 200 nM MMC or 5 & 10 μM BRD4770 for 24 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCA, anti-UBE2T, anti-USP1, anti-CHK1 pS345, anti-RPA, anti-RPA pS4/8, & anti-alpha -Tubulin antibodies. L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCD2; RBI, relative band intensity. (D) U2OS cells were incubated in the absence or presence of 2, 5, & 10 μM BRD4770 for 24, 48, or 72 h. Cells were fixed in ice-cold ethanol, stained with propidium iodide, & analyzed by flow cytometry. Cell cycle stage distributions were determined using FlowJo v10.2. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AICAR‐induced FANCD2 monoubiquitination is dependent on AMPK. (A) Inhibitor of AMPK blocks AICAR‐induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 μm of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 to visualize monoubiquitinated FANCD2 (Ub‐FANCD2). (B) AMPK inhibitor abrogates AICAR‐induced FANCD2 nuclear foci formation in GM00637I fibroblasts. GM006387I cells grown on coverslips were pretreated with 5 μm Compound C 1 h prior to 0.25 mm AICAR treatment for 24 h. FANCD2 foci were visualized by immunofluorescence staining & confocal microscopy. Representative images are shown at the top. The number of foci per cell was counted & plotted for ≥ 20 cells (bottom panel). The values represent the mean ± SEM (Student's t‐test, *P < 0.05; ***P < 0.001). (C) Knockdown of AMPK alpha 1 inhibits AICAR‐induced FANCD2 monoubiquitination in Caki‐1 cells. Caki‐1 cells were transfected with siRNAs (siControl or siAMPK#8) & after 48 h, AICAR was treated for 24 h. FANCD2 monoubiquitination was monitored by immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - LNK depletion restores FA-like human progenitor cells. a depicts a schematic overview of isolation of primary human HSPCs for lentiviral transduction followed by CFC assay. b UCB-derived CD34 + cells were sequentially infected with lentiviruses expressing shRNA to Luciferase (Luc) or FANCD2 (D2) with GFP marker, followed by shRNA to Luc or LNK with mCherry marker. GFP + mCherry + cells were then sorted & plated onto semi-solid methylcellulose media. Colony forming progenitor numbers are shown with a two-tailed Student's t-test. Representative data from two independent repeats are shown. c TF-1 cells with shRNA-mediated depletion of LNK or FANCD2 or CRISPR/Cas9-mediated depletion of LNK were analyzed for depletion efficiency by western blot. Luc, luciferase. gRNA, guide RNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30254368), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - SAN1 functions independently of the FA pathway & does not affect FA pathway activation. a, b CSAs of HeLa WT & SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin & MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT & SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells & SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT & SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA & exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - SAN1 functions independently of the FA pathway & does not affect FA pathway activation. a, b CSAs of HeLa WT & SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin & MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT & SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells & SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT & SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA & exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - The FANCD2 NLS is required for the nuclear localization of a subset of FANCI.(A) FA-D2 patient cells or FA-D2 cells stably expressing FANCD2-WT were incubated in the absence (NT) or presence of MMC for 24 h, fixed, stained with rabbit polyclonal anti-FANCD2 or anti-FANCI antibody & counterstained with phalloidin & DAPI. AF-488, Alexa Fluor 488. (B) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, & FANCD2-3N were incubated in the absence (NT) or presence of MMC for 24 h, fixed, & stained with rabbit polyclonal anti-FANCI antibody, & counterstained with phalloidin & DAPI. At least 300 cells were scored for cytoplasmic (Cyto.), nuclear (Nucl.), & both cytoplasmic & nuclear (Both) localization of FANCI. (C) COS-7 cells were transiently transfected with no DNA, FANCI-GFP, FANCI-GFP plus FANCD2-V5-WT, or FANCI-GFP plus FANCD2-V5-∆N57. Whole-cell lysates were immunoprecipitated with anti-V5 or anti-GFP antibodies & immune complexes immunoblotted with anti-GFP & anti-V5 antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts & FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, & FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FAN1 PIP-box motif is not required for FAN1 foci formation upon exposure to MMC. a Total cell extracts & chromatin-enriched fractions of U2OS cells expressing the indicated eGFP-FAN1 variants, treated or mock-treated with MMC (150 ng/ml, 24 h), were analysed by immunoblotting using the indicated antibodies. A representative blot of three independent experiments is shown. b Cells as in a were immunostained with anti-FANCD2 antibody. Representative images are shown. Scale bar: 25 μm. c, d Quantification of eGFP-FAN1 foci count (c) & the sum of their intensities (d) was obtained from QIBC analysis of b. Median levels are indicated by black bars. Statistical analyses were carried out using unpaired, two-tailed t-tests. P values expressed as ***(P < 0.01) were considered significant, n = 3. e Total cell extracts derived from cells as in a, treated or mock-treated with MMC (150 ng/ml, 24 h), were incubated with anti-eGFP affinity resin. Inputs & immunoprecipitates were analysed by immunoblotting with the indicated antibodies. A representative blot of two independent experiments is shown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29051491), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts & FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, & FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - DNA damage & cell survivalas a function of FANCD2 status. (a) Representative images of the formation of gamma H2AX foci in PD20 versus wild-typecomplemented cells 30 minutes after completion of H2O2 treatment. (b) Quantification of gamma H2AX foci response. Data represent means with upper standard error based on two independent experiments. (c) G2-type chromosomal aberrations are expressed as breaks per cell as a function of increasing H2O2 concentration in FANCD2-deficient andwild-type complemented PD20 cells. Data represent means with SEM based on at least three repeat experiments. (d) Apoptosis induction by H2O2 (50 μM) in cells with or without wild-type FANCD2 using fluorescence microscopy to assess apoptotic morphology by DAPI staining & flow cytometric analysis for sub-G1 DNA content. Representative experiments based on theapoptotic response at 24 hours are shown (similar results were obtained at 48 hours & with 25 μM H2O2). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18483568), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FANCD2 NLS is required for efficient FANCD2 & FANCI monoubiquitination & chromatin association.(A) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-K561R, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were incubated in the absence & presence of 250 nM MMC for 18 h, & whole-cell lysates were immunoblotted with antibodies to FANCD2, V5, FANCI & alpha -tubulin. The FANCD2 & FANCI L/S ratios are the ratios of monoubiquitinated to nonubiquitinated protein, & were calculated by measuring protein band intensities using ImageJ image processing & analysis software (//rsb.info.nih.gov/ij/). (B & C) FA-D2 cells stably expressing FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were treated as above & cell pellets were fractionated into soluble (S) & chromatin-associated (C) fractions. Fractions were immunoblotted with antibodies against V5, FANCI, alpha -tubulin & H2A. W, unfractionated whole cell extract. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AICAR‐induced FANCD2 monoubiquitination requires FANCA. (A) AICAR‐induced FANCD2 monoubiquitination does not occur in transformed FANCA−/− fibroblasts originated from a patient with defective FANCA. Transformed FANCA−/− (GM06914B) or normal fibroblasts (GM00637I) were treated with 0.25 mm AICAR for 24 h, & monoubiquitinated FANCD2 (Ub‐FANCD2) was visualized by immunoblotting. (B) AICAR‐induced FANCD2 nuclear foci formation is abrogated in FANCA−/− fibroblasts. FANCD2 nuclear foci were visualized by immunofluorescence staining & confocal microscopy. (C) Knockdown of FANCA abolishes FANCD2 monoubiquitination after AICAR treatment in Caki‐1 cells. Caki‐1 cells were transfected with FANCA‐targeting siRNAs (siFANCA#5 or siFANCA#6), treated with AICAR & subjected to FANCD2 immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Acetaldehyde toxicity to human FANCD2‐deleted human cells & FANCD2 ubiquitylation in BRCA2‐deleted cellsA, BHuman DLD1 cells in which FANCD2 was deleted with CRISPR/Cas9 & control cells were incubated with the indicated concentrations of cisplatin (A) or acetaldehyde (B) for 6 days before processing for dose‐dependent viability assays. Graphs are representative of two independent experiments, each performed in triplicate. Error bars represent SD of triplicate values obtained from a single experiment. Inset, Western blot detection of FANCD2 expression. SMC1 was used as a loading control.CBRCA2‐proficient (+BRCA2) or BRCA2‐deficient (−BRCA2) DLD1 cells were incubated with 4 mM acetaldehyde for 48 h before being processed for immunoblotting as indicated.DH1299 cells expressing a DOX‐inducible BRCA2 shRNA were grown in the presence or absence of DOX & transfected with control or FANCD2 siRNA before being processed for immunoblotting as indicated. DOX, doxycycline. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28729482), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - BRCA2-deficient cells accumulate single-stranded DNA lesions in G2. a gamma H2AX foci analysis in early mitotic cells. Cells were untreated, or treated with RO-3306 (10 µM, 24 h) to delay mitotic entry, & released for 1 h before analysis of gamma H2AX & FANCD2 foci pairs in early mitotic cells. Representative images are shown (left). Analysis is by an unpaired two-tailed t-test. n = 3. Scale bars 10 µm. b–g Cells treated with the indicated siRNAs or mirin (50 µM, 5 h) were incubated with EdU for 30 min & then analyzed for gamma H2AX & RPA foci in G2 cells (EdU–, 2N DNA content). Representative deconvolved images are shown (b). Quantification of gamma H2AX foci (top, c–g) & RPA+ gamma H2AX foci (bottom, c–g) for BRCA2-deficient cells (c), cells transfected with siRNAs (d, RAD51; e SMARCAL1; f EXO1 & DNA2), & cells treated with mirin (g) are shown. n ≥ 3. Scale bars 10 µm. h, i Cells were incubated with EdU as in b before analysis of gamma H2AX & pCHK2-T68 (h) or pATM-S1981 (i) foci in G2 cells. Representative deconvolved images with magnified inset highlighting foci co-localization are shown (left in each panel). n ≥ 3. Scale bars 10 µm. Error bars s.d. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 (unpaired two-tailed t-test) Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-00634-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - BRCA2 deficiency causes DNA under replication that results in abnormal mitoses. a–cBRCA2∆Ex3-4/– cells released for 22 h from serum starvation to increase mitotic cells, incubated w/ EdU for 1 h, & then analyzed for mitotic DNA synthesis. Early mitotic cells defined as being in prophase, prometaphase,/metaphase analyzed for EdU foci that co-localize w/ FANCD2 foci pairs. a Representative images of mitotic DNA synthesis. Scale bars 10 µm. b Percent early mitotic cells containing EdU foci, analyzed by an unpaired two-tailed t-test. n = 3. c FANCD2 foci pairs w//w/out EdU foci co-localization. Graphs represent pooled results of 3 independent experiments, each analyzed by a two-tailed Mann–Whitney test. Median FANCD2 foci pair number, red bars. Image collected & cropped by CiteAb from following publication (//www.nature.com/articles/s41467-017-00634-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FANCD2 NLS is required for efficient FANCD2 & FANCI monoubiquitination & chromatin association.(A) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-K561R, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were incubated in the absence & presence of 250 nM MMC for 18 h, & whole-cell lysates were immunoblotted with antibodies to FANCD2, V5, FANCI & alpha -tubulin. The FANCD2 & FANCI L/S ratios are the ratios of monoubiquitinated to nonubiquitinated protein, & were calculated by measuring protein band intensities using ImageJ image processing & analysis software (//rsb.info.nih.gov/ij/). (B & C) FA-D2 cells stably expressing FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were treated as above & cell pellets were fractionated into soluble (S) & chromatin-associated (C) fractions. Fractions were immunoblotted with antibodies against V5, FANCI, alpha -tubulin & H2A. W, unfractionated whole cell extract. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Detection of FANCD2 foci formation in human lung tumors by the FATSI staining analysis. The paraffin-embedded lung tumor tissues sections were deparaffinized & rehydrated. The tissue sections were incubated with a primary antibody cocktail of rabbit polyclonal FANCD2 antibody (Novus Biologicals, Littleton, CO, USA) at a dilution of 1:1000 & a monoclonal anti-Ki67 mouse antibody (Dako, Carpenteria, CA, USA) at a dilution of 1:150 for 1 h at room temperature. Sections then were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG & Alexafluor 594 donkey anti-mouse secondary for 1 h at room temperature. The sections were mounted on glass slides using a 4′ 6-diamidino-2-phenylindole (DAPI)-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA). The slides were analyzed under a fluorescence microscope. (A) FANCD2 foci positive NSCL tumor, & (B) FANCD2 foci negative NSCL tumor. Magnification: 1000×. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Normal DNA damage responses in FANCD2-deficient cells treated with hydrogen peroxide (H2O2). (a) Illustration of RAD51 foci formation in PD20 & PD20-wtD2 cells 5 hours after exposure to MMC (0.25 μg/mL for 1 hour) or H2O2 (25 μM for 2 hours). (b) Induction of RAD51 foci formation above background levels by MMC or H2O2. Because equal drug concentrations were used, rather than isoeffective concentrations with regard to cell survival, the extent of foci induction between FANCD-deficient & -complemented cells is not directly comparable. Data represent means with upperstandard error based on three independent repeats. (c) DNA synthesis measured by BrdU pulse labeling in cells treated with ionizing radiation (IR, 8 Gy) or H2O2 (25 μM). Data represent means with upper standard error based on twoindependent experiments. (d) Cell cycle distribution of propidium-iodide cell populations by flow cytometry. A representative experiment is shown. Percentages of cells in the G1, S, & G2 (and M) phases of the cell cycle are indicated. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18483568), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Dicer inhibition prevents FANCD2 foci formation after replication stress without affecting FANCD2 mono-ubiquitylation.(A) HCT116 cells were treated with control or Dicer siRNA & then with aphidicolin to induce replicative stress. They were then stained for FANCD2 & analyzed by immunofluorescence microscopy. (B) Quantification of the experiments in panel (A) showing the percentage of FANCD2 positive cells. Error bars represent the SD of three independent experiments. To score the FANCD2 positive cells a threshold value was calculated on the base of the average number of foci in control cells for each replicate, using Image J. Unpaired t-test: *p < 0.05. (C) FANCD2 foci formation in WT & Dicer Ex5/Ex5 cells after replication stress induced by aphidicolin. (D) Quantification of the experiments in panel (C) showing the percentage of FANCD2 positive cells. Error bars represent the SD of three independent experiments. Unpaired t-test: **p < 0.005. (E) Western blotting showing the levels of FANCD2 mono-ubiquitylation (L-FANCD2) after siRNA-mediated inhibition of Dicer, in the presence or absence of replication stress induced by aphidicolin. Vinculin was used as a loading control. (F) Western blotting showing FANCD2 mono-ubiquitylation levels after Drosha inhibition by siRNA. (G) FANCD2 mono-ubiquitylation levels in WT & Dicer Ex5/Ex5 cells assayed by western blotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31320994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FAN1 PIP-box motif is not required for FAN1 foci formation upon exposure to MMC. a Total cell extracts & chromatin-enriched fractions of U2OS cells expressing the indicated eGFP-FAN1 variants, treated or mock-treated with MMC (150 ng/ml, 24 h), were analysed by immunoblotting using the indicated antibodies. A representative blot of three independent experiments is shown. b Cells as in a were immunostained with anti-FANCD2 antibody. Representative images are shown. Scale bar: 25 μm. c, d Quantification of eGFP-FAN1 foci count (c) & the sum of their intensities (d) was obtained from QIBC analysis of b. Median levels are indicated by black bars. Statistical analyses were carried out using unpaired, two-tailed t-tests. P values expressed as ***(P < 0.01) were considered significant, n = 3. e Total cell extracts derived from cells as in a, treated or mock-treated with MMC (150 ng/ml, 24 h), were incubated with anti-eGFP affinity resin. Inputs & immunoprecipitates were analysed by immunoblotting with the indicated antibodies. A representative blot of two independent experiments is shown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29051491), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

An intracellular stain was performed on A431 human skin carcinoma cell line using Rabbit anti-FANCD2 Affinity Purified Polyclonal Antibody conjugated to DyLight 550 (Catalog # NB100-182R, blue histogram) or matched control antibody (Catalog # NBP2-24981R, orange histogram) at 2.5 µg/mL for 30 minutes at RT.

FANCD2 was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-FANCD2 Antigen Affinity Purified Polyclonal Antibody conjugated to Biotin (Catalog # NB100-182B) at 5 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Detection of FANCD2 foci formation in human lung tumors by the FATSI staining analysis. The paraffin-embedded lung tumor tissues sections were deparaffinized and rehydrated. The tissue sections were incubated with a primary antibody cocktail of rabbit polyclonal FANCD2 antibody (Novus Biologicals, Littleton, CO, USA) at a dilution of 1:1000 and a monoclonal anti-Ki67 mouse antibody (Dako, Carpenteria, CA, USA) at a dilution of 1:150 for 1 h at room temperature. Sections then were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG and Alexafluor 594 donkey anti-mouse secondary for 1 h at room temperature. The sections were mounted on glass slides using a 4′ 6-diamidino-2-phenylindole (DAPI)-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA). The slides were analyzed under a fluorescence microscope. (A) FANCD2 foci positive NSCL tumor, and (B) FANCD2 foci negative NSCL tumor. Magnification: 1000×. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25566506), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Av, Ca, Ma, Pm, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Av, Ca, Ma, Pm, Ze

Applications:

, , , , , , , , , , ,

Applications: , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB100-182AF405
Alexa Fluor 488	NB100-182AF488
Alexa Fluor 647	NB100-182AF647
Alexa Fluor 700	NB100-182AF700
Biotin	NB100-182B
DyLight 405	NB100-182V
DyLight 488	NB100-182G
DyLight 550	NB100-182R
DyLight 650	NB100-182C
FITC	NB100-182F
HRP	NB100-182H

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-182SS

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NB400-104

SR-BI Antibody - BSA Free

(5 Reviews)

(243 Publications)

0.1 ml

NB400-104

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - SR-BI antibody was tested in human fibroblast samples fixed in 4% PFA and permeabilized in PBS (0.2% Tween). Primary incubation overnight at 4C using a 1:100 dilution in PBS (0.1% Tween) with 1% BSA. Secondary antibody is anti-rabbit conjugate to Alexa Fluor 488. SR-BI is shown in green and nucleus in blue (Hoescht 33342 stain).

Flow Cytometry: SR-BI Antibody [NB400-104] - An intracellular stain was performed on HeLa cells with SR-BI antibody NB400-104AF488 (blue) and a matched isotype control NBP2-24893AF488 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: SR-BI Antibody [NB400-104] - Detection of SR-BI in rat H4IIE total cell lysates and plasma membrane proteins. Photo courtesy of product review by verified customer.

Immunohistochemistry: SR-BI Antibody [NB400-104] - Immunolocalization of SR-BI in adult mink testis using NB400-104. SR-BI labeling is visible at the surface and along the outline of the large vacuole. Photo courtesy of R.M. Pelletier, University of Montreal.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot 8310) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a specific membrane signal of SR-BI protein in the murine hepatocytes.

<b>Genetic Strategies Validation. </b>Western Blot: SR-BI Antibody [NB400-104] - HCV replication was increased by CD36. Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 u g CD36-HA plasmid or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - Staining of human adrenal cortex.

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-SR-BI at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot R-4) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated mainly a membranous signal of SR-BI protein in the murine hepatocytes.

Flow Cytometry: SR-BI Antibody [NB400-104] - Analysis of Huh7 and HepG2 cells using SR-B1 antibody NB400-104. Courtesy of Bruno Sainz, Jr., PhD, University of Illinois at Chicago.

ABCA1, ABCG1 & SR-BI protein expressions. a & b Hepatic protein expressions of ABCA1, ABCG1 & SR-BI were significantly decreased in COMT−/− mice at GD 18.5, compared to C57BL/6 J mice. Decreased hepatic ABCA1 expression was also observed at 10 days postpartum. ATI-5261 increased ABCA1 & ABCG1 expression in the liver at 10 days postpartum. c Placental protein expressions of ABCA1 & ABCG1 was reduced in COMT−/− mice, compared to C57BL/6 J mice. ATI-5261 treatment significantly increased ABCA1 levels in the placenta of COMT−/− mice. d Representative immunoblots of the corresponding proteins in the placenta with mouse RAW264.7 cell lysate included as positive control. Similar results were obtained when the experiment was repeated with lysates prepared from three batches of tissues. Data are presented as mean ± SEM. Groups (n = 8 in all groups) were compared using one-way ANOVA with post-hoc analysis (Tukey’s procedure). *, p 2000) (g, h, i, respectively). Bar graph shows the mean percentage of positively stained cells. Error bars depict standard deviations. ****p 0.3) by one-way ANOVA. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SR-BI Antibody - BSA Free [NB400-104] - Effects of 13-HODE & LA in the presence & absence of PPAR alpha & PPAR gamma selective antagonists on molecular markers of cholesterol homeostasis in RAW264.7 macrophages. RAW264.7 cells were pre-treated without or with the PPAR alpha selective antagonist GW6471 or the PPAR gamma selective antagonist GW9662 & subsequently treated without (vehicle control) or with 2.5 μmol/L 13-HODE or 100 μmol/L LA for 24 h. Afterwards, cells were lysed & subsequently processed for western blotting as described in the materials & methods section. A, Representative immunoblots specific for ABCA1, ABCG1, SR-BI, LXR alpha , & beta -actin which was used for normalization are shown. B, Bars represent data from densitometric analysis & are means ± SD from three independent experiments (n = 3). Data are expressed as percentage of protein concentration of vehicle control cells. Results from statistical analysis are indicated: Significant effects are denoted with superscript letters. Bars marked without a common superscript letter significantly differ (P 2000) (g, h, i, respectively). Bar graph shows the mean percentage of positively stained cells. Error bars depict standard deviations. ****p 0.3) by one-way ANOVA. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SR-BI Antibody - BSA Free [NB400-104] - Differentiation & characteristics of iHLCs. (a) Cell morphology & culture conditions at different stages of differentiation from iPSCs into iHLCs. Bright-field images were taken at the indicated time points (scale bar 100 µm). (b) Sequential expression & repression of transcription factors marking successful differentiation. Cells were immunostained with antibodies against Oct 3/4 (pluripotency marker), the endodermal transcription factor GATA-4, & the hepatic marker HNF4 alpha at day 0, 5, & 10 of differentiation (scale bars 20 µm). Shown are representative fluorescence & DIC images. (c) mRNA expression levels of different hepatocyte-specific factors in iHLCs, Huh7.5 cells, & primary human hepatocytes (PHHs) were analysed via qRT-PCR. Expression levels are displayed as fold over iPSCs normalised to GAPDH & 18S rRNA (mean ± SEM, n = 3–4, *p 0.9) by one-way ANOVA. §P < 0.02 by unpaired Student's t-test, relative to 0 h time point. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-104AF350
Alexa Fluor 405	NB400-104AF405
Alexa Fluor 488	NB400-104AF488
Alexa Fluor 532	NB400-104AF532
Alexa Fluor 594	NB400-104AF594
Alexa Fluor 647	NB400-104AF647
Alexa Fluor 700	NB400-104AF700
Alexa Fluor 750	NB400-104AF750
Alexa Fluor Plus 405	NB400-104AFP405
Alexa Fluor Plus 488	NB400-104AFP488
Alexa Fluor Plus 555	NB400-104AFP555
Alexa Fluor Plus 594	NB400-104AFP594
Alexa Fluor Plus 647	NB400-104AFP647
Alexa Fluor Plus 680	NB400-104AFP680
Allophycocyanin	NB400-104APC
Biotin	NB400-104B
CoraFluor 1	NB400-104CL1
DyLight 350	NB400-104UV
DyLight 405	NB400-104V
DyLight 488	NB400-104G
DyLight 550	NB400-104R
DyLight 594	NB400-104DL594
DyLight 650	NB400-104C
DyLight 680	NB400-104FR
DyLight 755	NB400-104IR
FITC	NB400-104F
HRP	NB400-104H
Janelia Fluor 525	NB400-104JF525
Janelia Fluor 549	NB400-104JF549
Janelia Fluor 646	NB400-104JF646
Janelia Fluor 669	NB400-104JF669
PE	NB400-104PE
PerCP	NB400-104PCP
mFluor Violet 450 SE	NB400-104MFV450
mFluor Violet 500 SE	NB400-104MFV500
mFluor Violet 610 SE	NB400-104MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-104SS

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NB100-143

Nbs1 Antibody - BSA Free

(8 Reviews)

(206 Publications)

0.05 ml

NB100-143

Immunocytochemistry/Immunofluorescence: Nbs1 Antibody [NB100-143] - ICC/IF analysis of Nbs1 in HeLa cells with Nbs1 Antibody [NB100-143]. Image from verified customer review.

Immunohistochemistry-Paraffin: Nbs1 Antibody [NB100-143] - FFPE human testis using Nbs1 antibody [NB100-143] at 1:200 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10 mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Staining was performed by Histowiz.

Western Blot: Nbs1 Antibody [NB100-143] - Analysis of HeLa whole cell lysate [NB800-PC1] using rabbit polyclonal NBS1 antibody [NB100-143]. Observed Molecular Weight at ~95 kDa.

<b>Genetic Strategies Validation. </b>Western Blot: Nbs1 Antibody [NB100-143] - Detection of NBS1 in Human Bone Osteosarcoma Epithelial Cells (U2OS whole cell lysate) using Nbs1 Antibody [NB100-143] at a dilution of 1:1000. Specificity was confirmed using a siRNA anti-NBS1 (targeting sequence: GGAAGAAACGUGAACUCAA). Image provided by Sebastien Britton of the Institute of Pharmacology and Structural Biology.

Western Blot: Nbs1 Antibody [NB100-143] - Western Blot with Nbs1 Antibody [NB100-143]. TIG-1 human primary fibroblasts, whole cell lysate (30 ug). Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Nbs1 Antibody [NB100-143] - Staining of HeLa cells using Nbs1 Antibody [NB100-143].

<b>Genetic Strategies Validation. </b>Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - TOPBP1 recruitment in response to rDNA breaks. TOPBP1 localization 2 h after I-Ppo1 transfection in NBS1-delta-N cells and NBS1-delta-N cells complemented with wild-type and mutant NBS1-mNG (percentage of cells with > 2 TOPBP1 caps are indicated; one of two experiments is shown). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31913317/) licensed under a CC-BY license.

HPV16 E2 partially colocalizes with the MRN complex proteins Mre11 and Nbs1. C33a cells were transfected with an HPV16 E2 expression plasmid, fixed in ice-cold methanol, and stained with an E2-specific antibody (green). DNA was stained with Hoechst 33342 (blue). (A) Endogenous Nbs1 was detected with an Nbs1-specific antibody (red/gray). (B) Endogenous Mre11 was detected with an Mre11-specific rabbit antibody (red/gray). Bar, 5 μm. Digital zoom is shown at the bottom.

Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - Replication centers of the integrated HPV recruit Mre11-Nbs1-Rad50 complex & Ku70/80 heterodimer.HeLa cells were transfected & analyzed as described previously. Co-immunostaining of HPV18 E1 (Alexa Fluor 568, second column) & the following proteins are presented: Mre11, Nbs1, Rad50, & Ku70/80 proteins (Alexa Fluor 488, first column). The localizations of the E1 & the respective DNA repair proteins are also shown in the third column as a merged image & DAPI stained nuclei are presented in the fourth column. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/19390600), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - Loss of MUS81 activates DNA damage responseA. MUS81 depleted U2OS cells accumulate NBS1 foci defining sites of DNA damage. B. Partial co-localization of NBS1 foci & RPA foci in MUS81-depleted cells. C. Co-localization of RPA foci to ssDNA regions. Cells were labelled with BrdU for 3 days concomitant with MUS81 depletion. BrdU was detected at ssDNA regions by immunofluorescence. D. NBS1 foci arise in cells that are both positive & negative for Cyclin A expression. Scale bars, 5 μm. E. NBS1 foci form largely at non-telomeric loci. F. Partial co-localization of NBS1 foci with PML nuclear bodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26415217), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Nbs1 Antibody [NB100-143] - HPV16 E2 partially colocalizes with the MRN complex proteins Mre11 & Nbs1. C33a cells were transfected with an HPV16 E2 expression plasmid, fixed in ice-cold methanol, & stained with an E2-specific antibody (green). DNA was stained with Hoechst 33342 (blue). (A) Endogenous Nbs1 was detected with an Nbs1-specific antibody (red/gray). (B) Endogenous Mre11 was detected with an Mre11-specific rabbit antibody (red/gray). Bar, 5 μm. Digital zoom is shown at the bottom. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28031358), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Nbs1 Antibody [NB100-143] - UFL1 regulates the ATM signaling. a U2OS cells expressing UFL1 tet-on shRNA1 were irradiated with 2 Gy IR. Thirty minutes later, cells were lysed & blotted with indicated antibodies. b Representative picture of gamma H2AX, 53BP1, & BRCA1 foci in control (Dox-) & UFL1 knockdown (Dox+) U2OS cells 1 h after 0.5 Gy treatment. Scale bars, 10 µm. c–e Quantification of intensities of gamma H2AX, 53BP1, & BRCA1 foci in control (Dox-) & UFL1 knockdown (Dox+) U2OS cells. Data presented as mean ± SD of n = 50 cells. **p 100 cells. Error bars = SD. *** p-value <0.0001, unpaired two-tailed t-test.DOI://dx.doi.org/10.7554/eLife.07270.005Inhibition of oxidative stress reduces cell migration and invasion.ROS inhibitor NAC reduces cell migration to a similar extent as ATM-depletion. siATM and siNC control MDA-MB-231 cells were either untreated or treated with NAC and analyzed as in Figure 2D for cell migration. Right panel: Western blot of ATM depletion and quantification of cell migration for each sample.DOI://dx.doi.org/10.7554/eLife.07270.006 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26030852), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Ha, Ma, Pm

Species Glossary

Reactivity: Hu, Mu, Ha, Ma, Pm

Applications:

, , , , , , , , , , , ,

Applications: , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-143SS

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NB500-170

Ki67/MKI67 Antibody - BSA Free

(8 Reviews)

(196 Publications)

0.1 ml

NB500-170

Immunohistochemistry: Ki67/MKI67 Antibody [NB500-170] - Senescent MSPCs are characterized by loss of nestin expression. Double-immunofluorescence images of femoral metaphysis sections from 4-, 6-, 8-, and 12-week-old male Nestin-GFP mice using antibodies against GFP (green) and Ki67 (red). DAPI stains nuclei blue. GP growth plate. PS primary spongiosa. Quantification of the percentage of GFP+ cells that express Ki67. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-01509-0) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody [NB500-170] - Ki-67/MKI67 Antibody [NB500-170] - Tissue section of human tonsil using 1:50 dilution of rabbit anti-KI67 antibody. The staining was developed with HRP labeled anti-rabbit IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Ki67 antibody generated a specific nuclear staining in the cells in germinal centers of the tested tonsil tissue.

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody [NB500-170] - Ki-67/MKI67 Antibody [NB500-170] - Human tonsil using 1:200 dilution of rabbit anti-KI67 antibody. The staining was developed with HRP labeled anti-rabbit IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Ki67 antibody generated a specific nuclear staining in the cells in germinal centers of the tested tonsil tissue.

Western Blot: Ki67/MKI67 Antibody - BSA Free [NB500-170] - Whole cell lysates from SUM159, MDA-MB-231 and MCF-7 cells were loaded with 30 ug/lane. 10% SDS-PAGE. Ki67/MKI67 antibody (NB500-170) was used for primary antibody: 1:2000, 4C, overnight. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody [NB500-170] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Ki67/MKI67 Antibody NB500-170 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: Ki67/MKI67 Antibody - BSA Free [NB500-170] - An intracellular stain was performed on U-251 MG cells with Ki67/MKI67 Antibody NB500-170AF594 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 594.

Western Blot: Ki67/MKI67 Antibody [NB500-170] - Ki-67/MKI67 Antibody [NB500-170] - Analysis of A431 (A) and Hek293 (B) cell lysate using Ki67 antibody (NB500-170) at 2 ug/ml.

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody [NB500-170] - Ki67 antibody was tested at 1:25 in HeLa cells with FITC (green). Nuclei and actin were counterstained with Dapi (blue) and Phalloidin (red).

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody [NB500-170] - Confocal immunofluorescent analysis of MCF7 cells using Ki67 antibody (NB500-170, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Immunocytochemistry/Immunofluorescence: Ki67/MKI67 Antibody [NB500-170] - A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB500-170 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody [NB500-170] - Ki-67/MKI67 Antibody [NB500-170] - Human tonsil.

Ki67/MKI67 was detected in immersion fixed MCF7 human breast cancer cell line using Rabbit anti-Ki67/MKI67 Affinity Purified Polyclonal Antibody conjugated to Biotin (Catalog # NB500-170B) at 5 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Ki67/MKI67 was detected in immersion fixed A431 human skin carcinoma cell line using Rabbit anti- Ki67/MKI67 Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB500-170AF488) (green) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

An intracellular stain was performed on A431 human skin carcinoma cell line using Rabbit anti-Ki67/MKI67 Affinity Purified Polyclonal Antibody conjugated to DyLight 650 (Catalog # NB500-170C, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.

Ki67/MKI67 was detected in immersion fixed U-251 MG human glioblastoma cell line using Rabbit anti- Ki67/MKI67 Affinity Purified Polyclonal Antibody conjugated to DyLight 650 (Catalog # NB500-170C) (light blue) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry: Ki67/MKI67 Antibody - BSA Free [NB500-170] - PKD2 enzymatic deficiency had no effect on the proliferation or apoptosis of intestinal epithelial cells in mice exposed to DSS.(a) Representative 200× immunohistochemical images (left) & quantification of Ki-67+ proliferating intestinal epithelial cells (right) in wild-type & PKD2SSAA/SSAA mice. Bar represents 5 μm. (b) Representative 200× immunohistochemical images (left) & quantification of cleaved Caspase-3+ apoptotic intestinal epithelial cells (right) in wild-type & PKD2SSAA/SSAA mice. Bar represents 5 μm. Results are representative of three separate experiments with three mice per group. All p > 0.05 as measured by one way-ANOVA among four groups. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34079), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Ki67/MKI67 Antibody - BSA Free [NB500-170] - Deletion of Vangl2, but not Vangl1, in the mammary gland results in narrow ducts & low cell turnover. (A) RT-qPCR analysis of Vangl1 & Vangl2 mRNA levels in the mammary glands of MMTV-Cre, MMTV-Cre;Vangl2flox/+ & MMTV-Cre;Vangl2flox/flox at 10 weeks of age. (B) Representative images of 10 week old, carmine stained mammary glands & quantification of total branch number from MMTV-Cre, MMTV-Cre;Vangl2flox/+ & MMTV-Cre;Vangl2flox/flox mice (n = 3 mice per genotype). (C) Histological analysis by H&E staining show normal duct formation in 10 week old MMTV-Cre & MMTV-Cre;Vangl2flox/+ glands, whereas ducts from MMTV-Cre;Vangl2flox/flox glands are narrow & have significantly diminished lumens (n = 3 mice/genotype). (D) Immunostaining in mammary ducts show normal distribution of luminal (Cytokeratin 8 (K8), green), & basal (K5), red; SMA magenta) cell populations. (E) Immunofluorescence of adhesion junctional protein E-Cadherin (CDH1, green) & apical membrane marker pERM (red). (F,G) Immunostaining in 10 week old MMTV-Cre;Vangl2+/+ & MMTV-Cre;Vangl2flox/flox glands & quantitation of (F) Ki67 & (G) Cleaved Caspase 3 (n = 3–5 mice per genotype). Lu indicates lumen. * indicates non-magnified duct. Data are represented as mean +/− SEM. Scale bar represents 100 μm (C,F,G) 50 μm (D) & 10 μm (E). Student’s t-test *p < 0.05 & ****p < 0.0001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31068622), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody - BSA Free [NB500-170] - Senescent MSPCs are characterized by loss of nestin expression. a, b Representative images of GFP immunofluorescence staining (green) & quantitative analysis of GFP+ cells in femoral primary spongiosa from 4, 6, 8, & 12-week-old male Nestin-GFP mice. Images in upper panels in a are lower power w/ boxes outlining area of higher power in lower panels. Numbers of GFP+ cells per mm2 tissue area in primary spongiosa (N. GFP+ cells/PS.Ar) b. 4W, 6W, 8W, & 12W represent 4-, 6-, 8-, & 12-week-old mice, respectively. DAPI stains nuclei blue. c, d Representative images of flow cytometry analysis (c) & % of CD45−GFP+ cells (d) in femoral metaphysis from 4-, 6-, 8-, & 12-week-old male Nestin-GFP mice. e, f Double-immunofluorescence images of femoral metaphysis sections from 4-, 6-, 8-, & 12-week-old male Nestin-GFP mice using antibodies against GFP (green) & Ki67 (red) e. DAPI stains nuclei blue. GP growth plate. PS primary spongiosa. Quantification of % of GFP+ cells that express Ki67 f. Five mice per group. Data are represented as mean ± s.e.m. *P < 0.05 as determined by ANOVA. g Diagram showing isolation of Nestin-GFP+ (red) & Nestin-GFP− (purple) mesenchymal stem/progenitor cells (MSPCs) by fluorescence-activated cell sorting. Detailed information on isolation of MSPCs from femoral metaphysis from Nestin-GFP mice are described in Supplementary Fig. 3 & Methods section. h–j The sorted cells cultured, & SA-beta Gal staining (h), BrdU incorporation (i), & p16INK4a immunostaining (j) performed, & representative images shown. k–m Quantification of % of cells that express SA-beta Gal (k), BrdU (l), & p16INK4a (m). n = 5. Data are represented as mean ± s.e.m. *P < 0.01 as determined by Student’s t-tests Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/29101351), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Ki67/MKI67 Antibody - BSA Free [NB500-170] - Cellular senescence occurs in primary spongiosa of long bone during late puberty. a–e Representative senescence-associated beta -galactosidase (SA-beta Gal) staining (blue) & quantitative analysis of SA-beta Gal+ cells in femoral metaphysis (a–c) & diaphysis (d, e) sections from increasing ages of male mice. 4, 6, 8, & 12W represent 4-, 6-, 8-, & 12-week-old mice, respectively. Images in a are lower power with boxes outlining the area of higher power in b. Numbers of SA-beta Gal+ cells per mm2 tissue area in primary spongiosa (N. SA-beta Gal+ cells/PS.Ar) (c) & diaphysis (N. SA-beta Gal+ cells/DP.Ar) (e). Counterstained with eosin (pink). f, g Representative images of immunofluorescence staining (f) & quantitative analysis of ki67+ (g) cells (red) in femoral primary spongiosa from 4, 6, 8, & 12-week-old male mice. DAPI stains nuclei blue. Images in upper panels in f are lower power with boxes outlining the area of higher power in lower panels. Five mice per group. Data are represented as mean ± s.e.m. Ar tissue area, DP diaphysis, GP growth plate, N number, PS primary spongiosa. *P < 0.01 as determined by ANOVA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29101351), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Ki67/MKI67 was detected in immersion fixed A431 human skin carcinoma cell line using Rabbit anti-Ki67/MKI67 Antigen Affinity Purified Polyclonal Antibody conjugated to Biotin (Catalog # NB500-170B) at 2 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Reactivity:

Hu, Mu, Rt, Po, Av

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB500-170AF350
Alexa Fluor 405	NB500-170AF405
Alexa Fluor 488	NB500-170AF488
Alexa Fluor 532	NB500-170AF532
Alexa Fluor 594	NB500-170AF594
Alexa Fluor 647	NB500-170AF647
Alexa Fluor 700	NB500-170AF700
Alexa Fluor 750	NB500-170AF750
Alexa Fluor Plus 405	NB500-170AFP405
Alexa Fluor Plus 488	NB500-170AFP488
Alexa Fluor Plus 555	NB500-170AFP555
Alexa Fluor Plus 594	NB500-170AFP594
Alexa Fluor Plus 647	NB500-170AFP647
Alexa Fluor Plus 680	NB500-170AFP680
Allophycocyanin	NB500-170APC
Biotin	NB500-170B
CoraFluor 1	NB500-170CL1
DyLight 350	NB500-170UV
DyLight 405	NB500-170V
DyLight 488	NB500-170G
DyLight 550	NB500-170R
DyLight 594	NB500-170DL594
DyLight 650	NB500-170C
DyLight 680	NB500-170FR
DyLight 755	NB500-170IR
FITC	NB500-170F
HRP	NB500-170H
Janelia Fluor 525	NB500-170JF525
Janelia Fluor 549	NB500-170JF549
Janelia Fluor 646	NB500-170JF646
Janelia Fluor 669	NB500-170JF669
PE	NB500-170PE
PerCP	NB500-170PCP
mFluor Violet 450 SE	NB500-170MFV450
mFluor Violet 500 SE	NB500-170MFV500
mFluor Violet 610 SE	NB500-170MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB500-170SS

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(select up to 3 products from this page to compare)

NB100-305

53BP1 Antibody - BSA Free

(6 Reviews)

(152 Publications)

0.1 mg

NB100-305

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-305] - 53BP1 Antibody [NB100-305] - Exosomes modulate the repair of DNA DSBs in irradiated recipient cells. Representative images of 53BP1 foci in BHY cells 6 hours after 2 Gy and transfer of BHY exosomes isolated 24 hours after irradiation with 0, 3, 6 or 9 Gy (53BP1 foci green, nuclei blue). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0152213), licensed under a CC-BY license.

Immunohistochemistry: 53BP1 Antibody [NB100-305] - Immunohistochemical staining of placental villi with 53BP1 Antibody (Catalog #NB100-305) at 40X magnification.

Western Blot: 53BP1 Antibody [NB100-305] - Western blot of whole cell lysate (20 ug/lane) from U2Os cells resolved on a 3 to 8 percent trisacetate gel. A band appears at approximately the theoretical molecular weight of 214 kDa.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - HCC1937 cells (human) were treated with PARPi for 96 h. Nuclei are stained in blue (DAPI) and 53BP1 antibody (foci in green). ICC/IF image submitted by a verified customer review.

Flow Cytometry: 53BP1 Antibody [NB100-305] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody NB100-305 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - Paraformaldehyde (4%) fixed Saos-2 cells (human osteosarcoma cell line) were stained with anti-53BP1 antibody followed by Alexa Fluor 488 secondary antibody. ICC/IF image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB100-305 at 2 ug/mL overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: 53BP1 Antibody [NB100-305] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-305AF488) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: 53BP1 Antibody - BSA Free [NB100-305] - DNA damage & DNA damage responses upon H2O2 challenge. a, c, e, g Confocal images showing the basal region of the organ of Corti cultures treated with either culture medium alone (a, e) or containing 0.5 mM H2O2 (c, g) for 5 h before being maintained in culture medium alone for 3 days. The samples were then immunolabeled for myosin 7A (red, a, c, e, g), gamma H2AX (green, a & c) & 53BP1 (green, e & g). Scale bars: a, c, e & g = 10 μm. b, d, f, h Higher magnification images of representative OHC & IHC nuclei from all conditions tested. Scale bar = 2.5 μm. i Representative Western blot analysis using antibodies against gamma H2AX, 53BP1, DDB2, p-Chk1, p-Chk2, p53, & beta -actin in whole cochlear extracts. j, k Histograms representing the levels of gamma H2AX, 53BP1, DDB2, p-Chk1, p-Chk2, & p53 in control & in 0.4 & 0.5 mM H2O2-exposed groups (n = 6 cochleae per condition). beta -Actin served as a loading control. Data are expressed as mean ± SEM. One-way ANOVA test followed by post hoc Tukey’s test (*P ≤ 0.04, **P ≤ 0.01, ***P ≤ 0.001 vs. H2O2 0 mM). All experiments were performed in triplicate Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30693443), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-305] - Assessment of satellite cell radiosensitivity & DNA repair pathways w/ age.(A–B) Expression of major proteins involved in homologous recombination & non-homologous end joining detected using WB. Protein nuclear extracts prepared from activated satellite cells 72 hrs post muscle injury. Lamin B used as loading control. (A) Representative images of WBs. (B) Quantification using Image J software. Data represents mean +/− SEM, n = 3 to 5 (n = a pool of muscles from 3 mice), 2-tailed unpaired Student's t-test, no significant difference. (C) Graphical representation of data from Table S1. Gene expression of old relative to young satellite cells presented as fold change w/ a 95 % confidence interval for the 12 genes w/ a statistically significant relative expression, n = 3 mice per group, two-tailed unpaired Student's t-test, *: p value 0.05. Shown are the mean +/-SEM for three independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27494840), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$The production of DSBs depends on Top1 degradation in CPT-treated quiescent cells. (A–C) Serum-starved WI38 hTERT cells were co-transfected with siRNAs against cullin 3 and cullin 4B or against a control sequence and then treated with DMSO (−CPT) or 25 μM CPT (+CPT) for 1 h. (A and B) Western blotting of the indicated proteins. alpha Tubulin: loading control. (C) Number of gamma H2AX foci per nucleus from one representative experiment (246–348 nuclei were analyzed for each treatment) out of three. ***P 5 53BP1 foci/nucleus in a single plane of view) in UM-SCC-6 (C) and UT-SCC-5 (D) cells. Shown are mean percentages of three biological replicates. Immunofluorescent representative images of examples of UM-SCC-6 or UT-SCC-5 cells, 24 h post irradiation, with or without 5 uM BMS303141 pre-treatment. Nuclei in blue, 53BP1 foci in red. (E) Homologous recombination assay. U2OS cells have been stably transfected with a construct containing a GFP gene containing a I-SceI restriction site (SceGFP) as well as a stop codon at the same site. Upon transfection with an I-SceI encoded plasmid, a double strand break is induced at the I-SceI site. Repair of this break via homologous recombination occurs through utilization of the internal GFP (iGFP) fragment downstream of the SceGFP. Successful homologous recombination results in the restoration of a functional GFP gene. (F) U2OS DRGFP cells were treated for 6 h with 5 uM BMS303141 or 5 uM BO2 (Rad51 inhibitor), and transfected with pCBASceI plasmid. 48 h post transfection, cells were harvested and the GFP positive fraction was determined using flow cytometry. Representative images from flow cytometry experiments are shown on the right, with DMSO Sce- as a negative control, and 5 uM B02 Sce+ as a positive control. In the graph, data are shown from two biological replicates. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31817870), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

53BP1/RIF1 work epistatically to counteract BRCA1/CtIP-mediated end resection to regulate DSB repair pathway choiceA. HeLa cells harboring pGC were treated with the indicated siRNAs before transfection with I-SceI-expressing vector to induce DSBs. After 48h, the percentage of GFP+ cells was measured as an indication for HR efficiency. B.-D. Asynchronous A549 cells were treated with the indicated siRNAs before irradiation with 2Gy and CtIP B. and RPA foci C. were monitored at 2h while RAD51 D. foci were enumerated at 4h. E. Left panel: representative photos for the colocalization between 53BP1 and RIF1 foci in A549 cells at 2h after 2Gy. Right panel: quantitation of 53BP1, RIF1 or colocalized foci in the absence (DMSO) or presence of ATM inhibitor (ATMi). F. Left panel: representative micrographs for 53BP1 or RIF1 foci in irradiated cells after depletion of either proteins. Right panel: quantitation of experiments presented in the left panel. At least 100 nuclei were counted. In all cases, the number of foci measured in non-irradiated cells was subtracted (relative). Shown are the mean +/-SEM for three independent experiments. sc: scrambled RNA, si53: si53BP1, siR: siRIF1, siB: siBRCA1, and siC: siCtIP. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27494840), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

RAP1 protects telomeres against DNA damageWestern blotting showing the expression of RAP1 and TRF2 in MRC‐5 cells of different population doublings (PD). Senescent cells (PD 72) were left in culture for at least 4 weeks before harvesting for analysis.ChIP analysis of young (PD 30) and senescent (PD 72) MRC‐5 cells performed with either anti‐RAP1, anti‐TRF2, or an IgG antibody. The immunoprecipitated and input products were loaded into a slot blot membrane and hybridized with a telomere probe, and stripped and hybridized to an Alu probe in order to determine unspecific binding. Quantification was performed by normalizing the telomere signal of the immunoprecipitated to that of the input. Data represent mean +/- SD of three biological replicates.Immunofluorescence detection of 53BP1 (red) and a FISH probe staining telomeres (green) in young (PD 26), pre‐senescent (PD 66), and senescent (PD 72 + 4 weeks) MRC‐5 fibroblasts. The upper graph shows the percentage of telomere co‐localizing with 53BP1 (TIFs). Total DNA damage was measured by immunofluorescence detection of 53BP1. The lower graph shows the total number of 53BP1 foci per nucleus. Approximately 40–50 cells were analyzed per replicate and per condition. Bars represent SEM of two biological replicates.Data information: Statistical analyses were performed using Mann–Whitney U‐test (*P < 0.05; **P < 0.001; ***P < 0.0001). Scale bar = 10 μm.Source data are available online for this figure. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32096305), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bt, Bv, Ca, Nm, Po, Pm, Eq, Fe, Ha, Pm-Or, Pm, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Bt, Bv, Ca, Nm, Po, Pm, Eq, Fe, Ha, Pm-Or, Pm, Rb, Sh

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-305AF350
Alexa Fluor 405	NB100-305AF405
Alexa Fluor 488	NB100-305AF488
Alexa Fluor 532	NB100-305AF532
Alexa Fluor 594	NB100-305AF594
Alexa Fluor 647	NB100-305AF647
Alexa Fluor 700	NB100-305AF700
Alexa Fluor 750	NB100-305AF750
Alexa Fluor Plus 405	NB100-305AFP405
Alexa Fluor Plus 488	NB100-305AFP488
Alexa Fluor Plus 555	NB100-305AFP555
Alexa Fluor Plus 594	NB100-305AFP594
Alexa Fluor Plus 647	NB100-305AFP647
Alexa Fluor Plus 680	NB100-305AFP680
Allophycocyanin	NB100-305APC
Biotin	NB100-305B
CoraFluor 1	NB100-305CL1
DyLight 350	NB100-305UV
DyLight 405	NB100-305V
DyLight 488	NB100-305G
DyLight 550	NB100-305R
DyLight 594	NB100-305DL594
DyLight 650	NB100-305C
DyLight 680	NB100-305FR
DyLight 755	NB100-305IR
FITC	NB100-305F
HRP	NB100-305H
Janelia Fluor 525	NB100-305JF525
Janelia Fluor 549	NB100-305JF549
Janelia Fluor 646	NB100-305JF646
Janelia Fluor 669	NB100-305JF669
PE	NB100-305PE
PerCP	NB100-305PCP
mFluor Violet 450 SE	NB100-305MFV450
mFluor Violet 500 SE	NB100-305MFV500
mFluor Violet 610 SE	NB100-305MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-305SS

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NB100-904

53BP1 Antibody - BSA Free

(4 Reviews)

(138 Publications)

0.1 ml

NB100-904

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB100-904 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry: 53BP1 Antibody - BSA Free [NB100-904] - Staining of 53BP1 in human renal cancer cells using DAB with hematoxylin counterstain.

<b>Biological Strategies Validation. </b>Western Blot: 53BP1 Antibody - BSA Free [NB100-904] - Western blot analysis of untreated U2OS cell lysate blotted with 53BP1 Antibody. The observed molecular weight here is ~220 kDa, and the theoretical molecular weight is 214 kDa.

<b>Biological Strategies Validation. </b>53BP1 Antibody - BSA Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0023.jpg

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Immunofluorescence of 53BP1 Antibody in HeLa cells. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Immunofluorescence of 53BP1 Antibody in C33A cells. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with 53BP1 Antibody conjugated to Alexa Fluor 488 (Catalog #NB100-904AF488) at 10ug/mL for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody NB100-904 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on HeLa cells (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-904AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-904C) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Chromatin opening activates DDR via a mechanism that requires catalytically functional LOXL2 but can be independent of DNA damage. d gamma -H2AX & 53BP1 staining & foci quantification are shown after immunofluorescence w/ indicated antibodies in non-replicative conditions. Dot graphs indicate number of gamma -H2AX (left graph) & 53BP1 (right graph) foci in control & LOXL2 KD cells. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/31462706), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Integrin-associated proteins affected cell cycle & DNA double strand repairA. Western blot & B. densitometric analysis of p53-wildtype A172 & U87MG cells 48 h after PINCH1, ILK or ILKAP knockdown. Non-specific siRNA served as the control. C. Percentage of BrdU positivity as a marker for S-phase cells at different time points after siRNA transfection. Representative images are shown. D. Immunofluorescence staining of 53BP1 (green) & gamma H2AX (red) & E. quantitative analysis of foci numbers in irradiated A172 & U87MG cells after PINCH1, ILK or ILKAP depletion. Nuclei were stained with DAPI (blue). Data are mean ± SD (n = 3; t-test; *P 10 IR-induced foci. Means±S.D. are shown, n=3. *P-value200 cells were counted per experiment). Expression levels of BRCA1 & 53BP1 were assessed by Western Blot on whole cell extracts 4 days after doxycycline treatment. A representative picture is shown in (C). The migration positions of molecular weight standards (in kDa) are indicated on the left. (D) Quantification of 3 Western blot experiments. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123333), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - hWDR76 interaction network & subnuclear localization suggest conservation of Cmr1 function.(a) Domain organization of Cmr1 & human WDR76. Human WDR76 shares 29% sequence similarity with Cmr1 from S. cerevisiae. Filled boxes indicate WD40 repeats. NLS, putative nuclear localization signal. (b) Human WDR76 interacts with HELLS, SUGT1, XRCC5, XRCC6 & the CCT–TRiC complex. SILAC-labelled HeLa cells were transfected with GFP-WDR76 or empty vector. GFP-WDR76 & its interacting proteins were enriched using GFP-Trap resin. Proteins were resolved by SDS–PAGE & digested in-gel with trypsin. Peptides were analysed on a quadrupole Orbitrap mass spectrometer. The plot shows log(10) SILAC ratios of proteins associated with GFP-WDR76 compared with background. WDR76 is highlighted in red & several other interactors are also indicated. (c) Human WDR76 localizes into nuclear foci. Twenty-four hours after GFP-WDR76 transfection, U2OS cells were treated with 10 μM MG132 or 1.5 mM MMS for 2 h. Immunofluorescence analyses of 53BP1 were performed with anti-53BP1 antibody. DAPI was used to stain nuclei. Scale bar, 20 μm. (d) Human WDR76 does not co-localize with PCNA. Stably expressing RFP-PCNA U2OS cells were transfected with GFP-WDR76. 24 h after transfection, cells were fixed & stained with DAPI. Scale bar, 20 μm. (e) Model for the role of Cmr1 in promoting replication recovery. See Discussion for details. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25817432), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Overexpression of RNF8 restores repair of DNA double-strand breaks in splicing-deficient cells. (a) U2OS cells were treated with DMSO, pladienolide B or isoginkgetin for 16 h, irradiated (6 Gy, 1 h recovery) 1 h prior to termination of this treatment, & then subjected to western blotting for H2A, RNF8 & beta -actin. (b & c) U2OS cells were transfected with either GFP-RNF8 or GFP-Empty for 2 h, followed by addition of pladienolide B, isoginkgetin or DMSO, incubation for an additional 5 h, irradiation with 6 Gy, fixation 1 h later, & immunostaining for (b) conjugated ubiquitin (FK2 antibody) & (c) 53BP1. The numbers in white indicate the percentage of 100 transfected, i.e, green cells whose nuclei contained >10 IR-induced foci. Means±S.D. are shown, n=3. (d) U2OS cells were treated as above, except that fixation was performed 24 h after irradiation & the immunostaining was for gamma H2AX. Again, the white numbers indicate the percentage of 100 transfected, i.e, green cells whose nuclei contained >10 gamma H2AX foci. Means±S.D. are shown, n=3. (e) The efficiency of HR measured in direct repeated-GFP U2OS cells transfected with I-SceI in combination with either Flag-Empty or Flag-RNF8 for 24 h, followed by addition of DMSO or pladienolide B & incubation for another 24 h. Means±S.D. are shown, n=3. *P200 cells were counted per experiment). Expression levels of BRCA1 & 53BP1 were assessed by Western Blot on whole cell extracts 4 days after doxycycline treatment. A representative picture is shown in (C). The migration positions of molecular weight standards (in kDa) are indicated on the left. (D) Quantification of 3 Western blot experiments. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123333), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - WRAP53 beta plays an important role in recruitment of factors involved in HR & NHEJ to DNA breaks in A2780 cells. (a) A2780 cells were transfected with siControl or siWRAP53#2 oligonucleotides for 48 h; exposed to IR (6 Gy), & 1 h later immunostained for gamma H2AX, MDC1, RNF168, FK2 (recognizes conjugated ubiquitin), BRCA1, 53BP1 & RAD51. (b) Quantification of the results in (a), as the percentage of 200 cells counted in each experiment whose nuclei contained >10 IR-induced foci. The error bars depict the S.E.M.; n=3, **PLeft: Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 1 mg/ml. A specific band was detected for 53BP1 at approximately 353 kDa (as indicated) using both 4 µg/ml and 20 µg/ml of Rabbit Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # NB100-904) followed by HRP-conjugated Goat Anti-Rabbit Secondary Antibody (Catalog # 042-206). This experiment was conducted under reducing conditions and using the 66-440kDa separation system. Right: Simple Western electropherogram showing the same Rabbit Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # NB100-904) tested at 4 µg/ml (blue line) and 20 µg/ml (green line) in the HepG2 human hepatocellular carcinoma cell line.

Reactivity:

Hu, Mu, Other, Pm, Pm-Or, Pm

Species Glossary

Reactivity: Hu, Mu, Other, Pm, Pm-Or, Pm

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-904AF350
Alexa Fluor 405	NB100-904AF405
Alexa Fluor 488	NB100-904AF488
Alexa Fluor 532	NB100-904AF532
Alexa Fluor 594	NB100-904AF594
Alexa Fluor 647	NB100-904AF647
Alexa Fluor 700	NB100-904AF700
Alexa Fluor 750	NB100-904AF750
Alexa Fluor Plus 405	NB100-904AFP405
Alexa Fluor Plus 488	NB100-904AFP488
Alexa Fluor Plus 555	NB100-904AFP555
Alexa Fluor Plus 594	NB100-904AFP594
Alexa Fluor Plus 647	NB100-904AFP647
Alexa Fluor Plus 680	NB100-904AFP680
Allophycocyanin	NB100-904APC
Allophycocyanin/Cy7	NB100-904APCCY7
Biotin	NB100-904B
CoraFluor 1	NB100-904CL1
DyLight 350	NB100-904UV
DyLight 405	NB100-904V
DyLight 488	NB100-904G
DyLight 550	NB100-904R
DyLight 594	NB100-904DL594
DyLight 650	NB100-904C
DyLight 680	NB100-904FR
DyLight 755	NB100-904IR
FITC	NB100-904F
HRP	NB100-904H
Janelia Fluor 525	NB100-904JF525
Janelia Fluor 549	NB100-904JF549
Janelia Fluor 646	NB100-904JF646
Janelia Fluor 669	NB100-904JF669
PE	NB100-904PE
PE/Atto594	NB100-904PEATT594
PE/Cy5.5	NB100-904PECY55
PE/Cy7	NB100-904PECY7
PerCP	NB100-904PCP
mFluor Violet 450 SE	NB100-904MFV450
mFluor Violet 500 SE	NB100-904MFV500
mFluor Violet 610 SE	NB100-904MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-904SS

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NB110-89474

CD11b Antibody - BSA Free

(12 Reviews)

(130 Publications)

0.1 ml

NB110-89474

Western Blot: CD11b Antibody - BSA Free [NB110-89474] - Western Blot detection of CD11b in RAW 264.7 whole cell lysates using [NB110-89474].

Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free [NB110-89474] - Staining of mouse pancreas using [NB110-89474] at 1:400 dilution. Nuclei counterstained with 4',6-diamidino-2-phenylindole (DAPI) (blue). ICC/IF image submitted by a verified customer review.

Immunohistochemistry: CD11b Antibody - BSA Free [NB110-89474] - Glibenclamide and Abcc8-/- suppress immune cell infiltration in EAE. White matter of lumbar spinal cord sections from WT control (a), untreated pid-30 WT/EAE (b), glibenclamide-treated pid-30 WT/EAE (c), and pid-30 Abcc8-/-/EAE (d) mice, stained with H&E or immunolabeled for CD45 (leucocyte), CD3 (T cells), CD20 (B cells), or CD11b (macrophage/microglia), as indicated; original magnification, x200 (H&E) or x400 (all immunolabelings). eleft panel: percent of quadrants with inflammatory cells on H&E; four mice/group. efour right panels: Quantification of CD-45-, CD3-, CD20-, and CD11b-expressing cells in white matter; four mice/group; ##P < 0.01 with respect to WT control; **P < 0.01, and ***P < 0.001 with respect to WT/EAE; scale bars, 100 um Image collected and cropped by CiteAb from the following publication (//www.jneuroinflammation.com/content/12/1/210) licensed under a CC-BY license.

Flow (Cell Surface): CD11b Antibody - BSA Free [NB110-89474] - Surface staining of CD11b in CT26 colorectal carcinoma tumor model. Using Alexa Fluor 405 conjugated version of the antibody (NB110-89474AF405). Image from verified customer review.

Immunocytochemistry/Immunofluorescence: CD11b Antibody - BSA Free [NB110-89474] - CD11b Antibody [NB110-89474] - CD11b antibody [NB110-89474] was tested in Raw264.7 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with 4',6-diamidino-2-phenylindole (DAPI) (blue) and DyLight 550 (red).

Immunohistochemistry: CD11b Antibody - BSA Free [NB110-89474] - Population of CD11b + myeloid progenitor cells differentiate into SMA + stromal cells within tumors and in vitro. Representative image of CD11b + SMA + double positive cells within tumor stroma. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32731354/) licensed under a CC-BY license.

Immunohistochemistry: CD11b Antibody - BSA Free [NB110-89474] - Immunohistochemical analysis of CD11b in human renal cancer with [NB110-89474] using 3,3'-Diaminobenzidine (DAB) with hematoxylin counterstain.

Immunohistochemistry-Frozen: CD11b Antibody - BSA Free [NB110-89474] - Immunohistochemical analysis of frozen sections of mouse spleen using [NB110-89474]. IHC-Fr image submitted by a verified customer review.

Immunohistochemistry: CD11b Antibody - BSA Free [NB110-89474] - Representative photomicrographs of cross sections of uteri collected from wild type (WT) mice 24 h after intrauterine infusion with 100 ul of vehicle or 100 ug ultrapure LPS from E. coli O111:B4, and Tlr4-/- mice similarly infused intrauterine with vehicle or LPS. Cross sections were stained with haematoxylin and eosin, and immunostained using a CD11b antibody for granulocytes (red) and DAPI for cell nuclei (white), and images collected at 10X magnification sections (C, F, I, L; bars = 200 um). At least 4 sections per animal and at least 4 fields per section were examined. Only WT mice infused with LPS show evidence of inflammation and accumulation of granulocytes in the endometrium. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0012906) licensed under a CC-BY license.

Flow (Cell Surface): CD11b Antibody - BSA Free [NB110-89474] - CD11b Antibody [NB110-89474] - A surface stain was performed on Raw264.7 cells with DyLight 550-conjugated [NB110-89474R] (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 10 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

Flow (Cell Surface): CD11b Antibody - BSA Free [NB110-89474] - An intracellular stain was performed on THP-1 cells with CD11b NB110-89474 (blue) and a matched isotype control NBP2-24891 (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Simple Western: CD11b Antibody - BSA Free [NB110-89474] - Specific band for Cd11b in 1.0 mg/mL of Dentate Gyrus from Rat Brain. [NB110-89474] was used at a dilution of 1:50. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Simple Western image submitted by a verified customer review.

Dual RNAscope ISH-IHC: CD11b Antibody - BSA Free [NB110-89474] - Formalin-fixed paraffin-embedded tissue sections of human lymph node were probed for CD11b mRNA (ACD RNAScope Probe, catalog #555098; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog #NB110-89474) at 1:50 dilution with 1 hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes.

Dual RNAscope ISH-IHC: CD11b Antibody - BSA Free [NB110-89474] - CD14 mRNA (red) and CD11b protein (green) were detected in formalin-fixed paraffin-embedded tissue sections of human malignant lymph node. ACD's Integrated Co-Detection Workflow was performed using ACD RNAScope Probe Hs-CD14 and CD11b antibody at 1:200 dilution. Tissue was stained on Leica Bond RX using RNAscope (TM) 2.5 LS Reagent Kit-RED, BOND Polymer Refine Detection (DAB) and Hematoxylin, BOND Polymer Refine Red Detection and Hematoxylin and RNA scope (TM) 2.5 LS Green Accessory Pack. Tissue was counterstained with 50% hematoxylin (blue).

CD11b staining in IMhu. Cells were stained with antibodies against the CD11b surface antigen. (A) Panel A shows positive staining for CD11b and (B) panel B shows nuclear DAPI staining. Magnitude 40×.

Western Blot: CD11b Antibody - BSA Free [NB110-89474] - Immunodensities of (a) CD11b, (b) GFAP & (c) NF-kappa B (p65) proteins with representative immunoblots in striatum from Munc18-OE (n = 5) & wild-type (n = 5) mice. Bar graphs are ratios of optical densities of our proteins of interest to beta -actin (42 kDa band), expressed as immunoreactivity in percentage of mean value of the WT group (100%). No differences were detected between groups for any of the analyzed proteins. Right panels are representative immunoblots for target proteins & beta -actin which included Munc18-OE (OE) & wild-type (WT) mice samples. The molecular masses were estimated from referenced standards. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25069615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD11b Antibody - BSA Free [NB110-89474] - Comparison of Ki-67 or CD11b immunoreactivity of Aldara-treated dorsal skin samples of C57BL/6 mice using OP or MP. Nuclear proliferation marker Ki-67 immunostaining on OP (a) & MP (b) dorsal skin tissue samples at 400x magnification. Dermal dendritic cells were labelled with anti-CD11b antibody on OP (c) & MP samples (d) at 400x magnification. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30842501), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CD11b Antibody - BSA Free [NB110-89474] - CD11b staining in IMhu. Cells were stained with antibodies against the CD11b surface antigen. (A) Panel A shows positive staining for CD11b & (B) panel B shows nuclear DAPI staining. Magnitude 40×. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31096716), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Population of CD11b + myeloid progenitor cells differentiate into SMA + stromal cells within tumors and in vitro. (A) Representative image of red fluorescent protein (RFP) + stromal cells in tumor from CCR2-RFP heterozygous SCID mouse. (B) RFP + SMA + double positive cells within tumor stroma. (C) CD11b + SMA + double positive cells within tumor stroma. (D) CD45 + CD11b + CD34+ myeloid progenitor cells in the mammary gland at 1.5 (n = 5 empty vector (EV), 7 CCL2) and 2.5 weeks (n = 5 EV, 7 CCL2) post-transplantation were quantified by flow cytometry. (E) CD45 + CD11b + CD34 + myeloid progenitor cells in the bone marrow at 1.5 weeks (n = 6 EV, 7 CCL2) and 2.5 weeks (n = 3 mice/group) post-transplantation were quantified by flow cytometry. (F) Representative brightfield image of colony formed by CD45 + CD11b + CD34 + myeloid progenitor cells isolated using fluorescence-activated cell sorting (FACS). (G) Colonies in culture co-stained with SMA and collagen I. Statistical differences determined by Mann–Whitney U test. Magnification bars = 50 um. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32731354), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv, Ce, ChHa, Pm, RM

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ce, ChHa, Pm, RM

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB110-89474AF350
Alexa Fluor 405	NB110-89474AF405
Alexa Fluor 488	NB110-89474AF488
Alexa Fluor 532	NB110-89474AF532
Alexa Fluor 594	NB110-89474AF594
Alexa Fluor 647	NB110-89474AF647
Alexa Fluor 700	NB110-89474AF700
Alexa Fluor 750	NB110-89474AF750
Alexa Fluor Plus 405	NB110-89474AFP405
Alexa Fluor Plus 488	NB110-89474AFP488
Alexa Fluor Plus 555	NB110-89474AFP555
Alexa Fluor Plus 594	NB110-89474AFP594
Alexa Fluor Plus 647	NB110-89474AFP647
Alexa Fluor Plus 680	NB110-89474AFP680
Allophycocyanin	NB110-89474APC
Allophycocyanin/Cy7	NB110-89474APCCY7
Biotin	NB110-89474B
CoraFluor 1	NB110-89474CL1
DyLight 350	NB110-89474UV
DyLight 405	NB110-89474V
DyLight 488	NB110-89474G
DyLight 550	NB110-89474R
DyLight 594	NB110-89474DL594
DyLight 650	NB110-89474C
DyLight 680	NB110-89474FR
DyLight 755	NB110-89474IR
FITC	NB110-89474F
HRP	NB110-89474H
Janelia Fluor 525	NB110-89474JF525
Janelia Fluor 549	NB110-89474JF549
Janelia Fluor 646	NB110-89474JF646
Janelia Fluor 669	NB110-89474JF669
PE	NB110-89474PE
PE/Atto594	NB110-89474PEATT594
PE/Cy5.5	NB110-89474PECY55
PE/Cy7	NB110-89474PECY7
PerCP	NB110-89474PCP
mFluor Violet 450 SE	NB110-89474MFV450
mFluor Violet 500 SE	NB110-89474MFV500
mFluor Violet 610 SE	NB110-89474MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB110-89474SS

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NB100-309

ATM Antibody (2C1) - Azide and BSA Free

(3 Reviews)

(132 Publications)

100 ul

NB100-309

Western Blot: ATM Antibody (2C1) [NB100-309] - Analysis of ATM in U2OS sarcoma cells using ATM antibody (2C1) [NB100-309]. Theoretical molecular weight 351 kDa. Western blot image submitted by a verifeid customer review.

Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309] - Human Kidney (formalin-fixed, paraffin-embedded) stained with ATM antibody (2C1) [NB100-309] at 5 ug/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.

<b>Genetic Strategies Validation. </b>Western Blot: ATM Antibody (2C1) [NB100-309] - Non-transfected (-) and transfected (+) HeLa whole cell extracts (60 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody (2C1) [NB100-309] diluted at 1:500.

<b>Genetic Strategies Validation. </b>Western Blot: ATM Antibody (2C1) [NB100-309] - Methylated EDSBs may be repaired by an ATM-dependent pathway. Immunoblots of ATM and DNA-PKcs in ATM shRNA-transfected HeLa cells, using ATM antibody (2C1) [NB100-309]. GAPDH is included as a loading control. Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-9-70), licensed under a CC-BY license.

Immunoprecipitation: ATM Antibody (2C1) [NB100-309] - MDA-MB-231 whole cell lysates were immunoprecipitated with 2 ug ATM Antibody (NB100-309), followed by Western blot (primary antibody: NB100-309 at 1:1000, 4C overnight. Western blot image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Western Blot: ATM Antibody (2C1) [NB100-309] - MCELNs decreased the DNA damage of H9C2 cells after radiation. Western blot images of p-ATM (NB100-306) and ATM (NB100-309) in H9C2 cells after 48 h of culture with indicated treatment. IR (-/+): 0/16 Gy X-ray; MCELNs (-/+): 0/10 ug/mL. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35509278/) licensed under a CC-BY license.

Western Blot: ATM Antibody (2C1) [NB100-309] - Western blot analysis of the extent of knockdown MRE11 & ATM, & the lack of effect of Mirin activation of ATM. The extent of shRNA-mediated knockdown of MRE11 & ATM are shown for (A) clones GFP-7F1 & GFP-6D1 & (B) clones EDS-7F2 & EDS-6J8. The extent of knockdown was determined by analyzing the intensity of the ATM & MRE11 bands relative to the intensity of the loading control GAPDH bands (see Table 1). (C) The effect of Mirin on activation of ATM was determined by analysis of phosphorylation of ATM in response to ionizing radiation. Cultures treated with DMSO alone, 20 μM Mirin, or knockdown of ATM were analyzed by western blot 30 min after exposure to 10 Gy of ionizing radiation. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26209132), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody (2C1) [NB100-309] - Rac1 inhibition abolishes IR-induced activation of both ATM & ATR signaling. (A) MCF-7 cells were treated with/without 20-Gy IR in the presence or absence of 100 μM NSC23766 & incubated for 1 hour at 37°C before analysis. To assess the ATM kinase activity, ATM was immunoprecipitated from cell lysate by using anti-ATM antibody (2C1) & assayed for ATM activity by using p53 recombinant protein as substrate. To measure the Chk2 activity, Chk2 was immunoprecipitated from cell lysate by using B-4 anti-Chk2 antibody & assayed for Chk2 activity by using Cdc25C recombinant protein as substrate. As controls, ATM & Chk2 protein levels in the immunoprecipitates (IP-WB) as well as in cell lysates (WB) were assessed with immunoblotting. (B) ATR & Chk1 were immunoprecipitated from cell lysates by using N-19 anti-ATR & G-4 anti-Chk1 antibody, respectively. ATR activity was assayed by using p53 recombinant protein substrate, & Chk1 activity assayed by using Cdc25C recombinant protein substrate. As controls, ATR & Chk1 protein levels in the immunoprecipitates (IP-WB), as well as in cell lysates (WB) were assessed with immunoblotting. (C) MCF-7 cells were exposed to IR at the indicated doses in the presence or absence of NSC23766, incubated for 1 hour, & assessed for Chk1 & Chk2 activities. *Kinase assay does not contain Cdc25C substrate. (D) T47D & ZR-75-1 cells were exposed to 10-Gy IR in the presence or absence of 100 μM NSC23766, incubated for 1 hour, & analyzed for Chk1 & Chk2 activities. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3164), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody (2C1) [NB100-309] - Inhibition of Rac1 by N17Rac1 mutant or Rac1 siRNA diminishes IR-induced G2/M checkpoint activation. (A) MCF-7 cells were infected with Ad.N17Rac1 or Ad.Control for 24 hours & exposed to 15-Gy IR. Left panel: the cells were analyzed for DNA content 24 hours after IR. The result depicts the percentage of cells with 4N-DNA content & is shown as mean ± SD of quadruplicate samples. *P < 0.001 (n = 4), significant difference from the irradiated Ad.Control-infected cells. Right panel: Inset: at 15 minutes after IR, the infected cells were analyzed for Rac1 activities (Rac1-GTP) & protein levels (total Rac1). Bar graph: mitotic cells in the cell samples were analyzed 2 hours after IR. The result depicts the percentage of mitotic cells & is shown as mean ± SD of triplicate samples. **P = 0.002 (n = 3), significant difference from the irradiated Ad.Control-infected cells. (B) Upper panel: MCF-7 cells transfected with Rac1 siRNA (Rac1) or control siRNA (Control) were incubated for the indicated times & analyzed for protein levels of Rac1 & Actin. Lower panel: After 2-day incubation, the siRNA-transfected cells were exposed to IR, incubated for 24 hours, & assessed for DNA content. Results depict the percentage of cells with 4N-DNA content & represent the mean ± SD of three separate experiments in duplicate samples. *P < 0.001 (n = 6), significant difference from the irradiated Control-siRNA transfected cells. (C) After 2-day incubation, siRNA-transfected cells were treated with/without 20-Gy IR, incubated for 1 hour, & analyzed for ATM, ATR, Chk1, & Chk2 activities. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3164), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody (2C1) [NB100-309] - HeLa whole cell extract and nuclear extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody [2C1] (NB100-309) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Western Blot: ATM Antibody (2C1) [NB100-309] - Whole cell extract (30 ug) was separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody [2C1] (NB100-309) diluted at 1:1000.

Human Testis (formalin-fixed, paraffin-embedded) stained with ATM antibody at 5 ug/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.

Reactivity:

Hu, Mu, Rt, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Pm

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NB400-101

SR-BI Antibody - BSA Free

(1 Review)

(114 Publications)

0.1 ml

NB400-101

<b>Genetic Strategies Validation. </b>Immunohistochemistry: SR-BI Antibody [NB400-101] - Immunofluorescent analysis of SR-BI localization in liver of normal, Tie2-Scarb1 x Scarb1-KO, and Scarb1-KO mice. Liver sections from normal (b) and Tie2-Scarb1 x Scarb1-KO mice (c) were stained for SR-BI (red) and cytokeratin 8-18 (green).Merge image for normal mouse (b) demonstrates strong presence of SR-BI in hepatocytes (b, yellow signal) and absence of detectable level of SR-BI protein in liver of Tie2-Scarb1 x Scarb1-KO mice (c). In Scarb1-KO mice there was no detectable level of SR-BI protein (d, staining for SR-BI). Blue = DAPI. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2015/607120/) licensed under a CC-BY license.

Western Blot: SR-BI Antibody [NB400-101] - Detection of SR-BI in mouse liver lysate (20 ug) using NB 400-101. ECL detection 5 seconds.

Flow (Intracellular): SR-BI Antibody [NB400-101] - An intracellular stain was performed on HeLa cells with NB400-101AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

$<b>Genetic Strategies Validation. </b>Western Blot: SR-BI Antibody [NB400-101] - Western blot and Coomassie stain of membrane fractions isolated from livers of normal C57Bl/6N (lane 1), Scarb1-KO (lane 2), and LIV11-SCARB1 x Scarb1-KO (lane 3) mice. 20 ug of membrane protein was loaded into each lane. (a)Western blot (anti-SR-BI antibody). (b) Coomassie stain (loading control for Western blot) encompassing the same MW region as SR-BI. Aliquots from the same tube were loaded for the Western blot and for the Coomassie-stained gel, which shows comparable loading between the three samples. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2015/607120/) licensed under a CC-BY license.$

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-101] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-SR-BI at 2 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: SR-BI Antibody [NB400-101] - Western blot analysis of ABCA1 and SR-B1 proteins from WT and Osbpl8KO mouse liver. The blots were probed with anti-beta-actin as a loading control. Densitometric quantification of the Western blot data is shown on the right. The results were normalized against beta-actin. The data represents mean +/- s.e.m. (n=4). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0058856), licensed under a CC-BY license.

Immunocytochemistry/ Immunofluorescence: SR-BI Antibody - BSA Free [NB400-101] - SR-BI distribution in polarized EC SVEC4-10 cells. SR-BI localization is presented in (a) as x-y image & orthogonal (x-z & y-z) views. (b) & (c) represent rendering of SR-BI staining (red) throughout the cells. Nuclei: blue. In (b) & (c) basolateral surface is on the top & the bottom, respectively. These results demonstrate the presence of SR-BI on both the apical & the basolateral sides of EC. The size of scale bar on (a) is 5 μm & on (b) & (c) 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26504816), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: SR-BI Antibody - BSA Free [NB400-101] - Immunofluorescent analysis of SR-BI localization in aorta of Tie2-Scarb1 × Scarb1-KO & Scarb1-KO mice. Mouse aorta sections were stained for SR-BI (a & d, red) & CD31 (b & e, green). Colocalization of SR-BI & CD31 was presented on (c) & (f). Blue = DAPI. (a, b, & c) Immunohistological analysis of SR-BI localization in aortic sections of Tie2-Scarb1 × Scarb1-KO mice. Colocalization of SR-BI & CD31 was found in aortic endothelial cells. In several EC stained for SR-B1 red color was present on both apical & basolateral sides (a). (d, e, & f) Immunohistological analysis of SR-BI localization in aortic sections of Scarb1-KO mice. There is no red signal in aorta from Scarb1-KO mice. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26504816), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: SR-BI Antibody - BSA Free [NB400-101] - Immunofluorescent analysis of SR-BI localization in liver of normal, Tie2-Scarb1 × Scarb1-KO, & Scarb1-KO mice. Liver sections from normal (a, b) & Tie2-Scarb1 × Scarb1-KO mice (c) were stained for SR-BI (red) & cytokeratin 8–18 (green). Merge image for normal mouse (b) demonstrates strong presence of SR-BI in hepatocytes (b, yellow signal) & absence of detectable level of SR-BI protein in liver of Tie2-Scarb1 × Scarb1-KO mice (c). In Scarb1-KO mice there was no detectable level of SR-BI protein (d, staining for SR-BI). Blue = DAPI. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26504816), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: SR-BI Antibody - BSA Free [NB400-101] - Western blot & Coomassie stain of membrane fractions isolated from livers of normal C57Bl/6N (lane 1), Scarb1-KO (lane 2), & LIV11-SCARB1 × Scarb1-KO (lane 3) mice. 20 μg of membrane protein was loaded into each lane. (a) Western blot (anti-SR-BI antibody). (b) Coomassie stain (loading control for Western blot) encompassing the same MW region as SR-BI. Aliquots from the same tube were loaded for the Western blot & for the Coomassie-stained gel, which shows comparable loading between the three samples. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26504816), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: SR-BI Antibody - BSA Free [NB400-101] - Liver mRNA & protein expression analysis of chow-fed Osbpl8KO mice.A: qPCR analysis of the quantity of the mRNAs identified at the bottom in chow-fed KO females (open bars) & males (closed bars). The mRNAs were quantified using ribosomal protein 36B4 message as a housekeeping reference. The data are expressed relative to quantity in littermate WT animals of the same gender, & represent mean ± s.e.m. (n = 6; *p<0.05, **p<0.01, T-test). B: Western blot analysis of ABCA1 & SR-B1 proteins in WT & KO mouse liver. The blots were probed with anti-beta -actin as a loading control. Densitometric quantification of the Western blot data is shown on the right. The results were normalized against beta -actin. The data represents mean ± s.e.m. (n = 4). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0058856), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: SR-BI Antibody - BSA Free [NB400-101] - Western blot & Coomassie stain of membrane fractions isolated from livers of normal C57Bl/6N (lane 1), Scarb1-KO (lane 2), & LIV11-SCARB1 × Scarb1-KO (lane 3) mice. 20 μg of membrane protein was loaded into each lane. (a) Western blot (anti-SR-BI antibody). (b) Coomassie stain (loading control for Western blot) encompassing the same MW region as SR-BI. Aliquots from the same tube were loaded for the Western blot & for the Coomassie-stained gel, which shows comparable loading between the three samples. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26504816), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Bv, ChHa, Md, Pm, Sj

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, ChHa, Md, Pm, Sj

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-101AF350
Alexa Fluor 405	NB400-101AF405
Alexa Fluor 488	NB400-101AF488
Alexa Fluor 532	NB400-101AF532
Alexa Fluor 594	NB400-101AF594
Alexa Fluor 647	NB400-101AF647
Alexa Fluor 700	NB400-101AF700
Alexa Fluor 750	NB400-101AF750
Alexa Fluor Plus 405	NB400-101AFP405
Alexa Fluor Plus 488	NB400-101AFP488
Alexa Fluor Plus 555	NB400-101AFP555
Alexa Fluor Plus 594	NB400-101AFP594
Alexa Fluor Plus 647	NB400-101AFP647
Alexa Fluor Plus 680	NB400-101AFP680
Allophycocyanin	NB400-101APC
Biotin	NB400-101B
CoraFluor 1	NB400-101CL1
DyLight 350	NB400-101UV
DyLight 405	NB400-101V
DyLight 488	NB400-101G
DyLight 550	NB400-101R
DyLight 594	NB400-101DL594
DyLight 650	NB400-101C
DyLight 680	NB400-101FR
DyLight 755	NB400-101IR
FITC	NB400-101F
HRP	NB400-101H
Janelia Fluor 525	NB400-101JF525
Janelia Fluor 549	NB400-101JF549
Janelia Fluor 646	NB400-101JF646
Janelia Fluor 669	NB400-101JF669
PE	NB400-101PE
PerCP	NB400-101PCP
mFluor Violet 450 SE	NB400-101MFV450
mFluor Violet 500 SE	NB400-101MFV500
mFluor Violet 610 SE	NB400-101MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-101SS

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NB100-104

ATM Antibody - BSA Free

(2 Reviews)

(100 Publications)

0.1 ml

NB100-104

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: ATM Antibody [NB100-104] - GM00637 cells with KU55933 pretreatment (a) were exposed to 50 uM spermidine or CCCP, followed by immunofluorescence analyses of total and p-ATM on Ser-1981. The scale bar is 10 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep24700) licensed under a CC-BY license.

Flow Cytometry: ATM Antibody [NB100-104] - An intracellular stain was performed on HeLa cells with Dylight 550-conjugated ATM Antibody [NB100-104R] (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

<b>Biological Strategies Validation. </b>Western Blot: ATM Antibody [NB100-104] - Analysis of double strand break repair in MCF-7 monolayer and mammosphere populations. Expression of proteins involved in the NHEJ pathway of DSB repair in response to increasing doses of ionizing radiation. Lysates were prepared from unirradiated cells and from cells harvested one hour after exposure to 1 or 10-Gy 60Co I3-radiation and analyzed by immunoblotting with antibodies against several DSB repair proteins. Phospho-ATM and phospho-DNA-PKcs refer to phosphorylation of these proteins at Ser1981 and Ser2056, respectively. Actin served as a loading control. Image collected and cropped by Citeab from the following publication (Senescence evasion by MCF-7 human breast tumor-initiating cells. Breast Cancer Res (2010)) licensed under a CC-BY license.

Western Blot: ATM Antibody [NB100-104] - Detection of ATM in Raji whole cell lysate using [NB100-104]. Observed molecular weight ~300 kDa. Theoretical molecular weight 351 kDa.

Immunocytochemistry/Immunofluorescence: ATM Antibody [NB100-104] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.05% Triton X-100. Then cells were incubated with [NB100-104] at a 1:100 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: ATM Antibody [NB100-104] - Analysis of PE conjugate of ATM Antibody [NB100-104PE]. An intracellular stain was performed on HeLa cells with ATM antibody [NB100-104PE] (blue) and a matched isotype control [NBP2-24893PE] (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin.

Flow Cytometry: ATM Antibody [NB100-104] - An intracellular stain was performed on HepG2 cells with PE-conjugated ATM antibody [NB100-104PE] (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin.

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104] - ATM is activated by spermidine in GM00637 cells.GM00637 cells with or without KU55933 pretreatment (a,b), & GM05849 cells (c) were exposed to 50 μM spermidine or CCCP, followed by immunofluorescence analyses of total & p-ATM on Ser-1981. The scale bar is 10 μm. Ratios of cells expressing p-ATM Ser-1981 to cells expressing total ATM were presented (d). 20–45 cells/condition from three experiments were collected. Values are mean ± SD, *p < 0.05 vs. control. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep24700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody [NB100-104] - Depletion of SMARCAL1 induces DNA damage & checkpoint activation in iSML1 iPSCs. (A,B) The iSML1 iPSCs were cultured for 7 & 14 days in the presence of doxycycline (DOX) to induce SMARCAL1 downregulation & then immunostained. The graphs (top) show quantification of the number of gamma -H2AX-positive cells (A) or ATM-pSer1981-positive cells (B). Representative images from triplicate experiments are shown (bottom). (C) Immunoblot detection of the indicated DDR proteins in iSML1 iPSCs after 14 days of continuous treatment with DOX. Lamin B1 was used as the loading control. Data are mean±s.e.m. from three independent experiments. *P≤0.05, **P≤0.01, ***P≤0.001 (two-way ANOVA test). ns, not significant. Scale bars: 10 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31515241), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody [NB100-104] - Analysis of double strand break repair in MCF-7 monolayer & mammosphere populations. (a) Cells were exposed to 4 Gy 60Co gamma -radiation & the relative degree of double-strand breakage (DSB) was determined by the comet assay under neutral conditions immediately after exposure & at the times indicated after exposure. (b) The 'comets' (n of about 100) were categorized according to the NIH LISTSERV (Comet Assay Interest Group web site) in which type 1 comets display the least DNA damage & type 5 the most. The error bars represent the mean ± standard error of the mean in both panels. The comets of the unirradiated cells are labeled Cont. (c) Expression of proteins involved in the NHEJ pathway of DSB repair in response to increasing doses of ionizing radiation. Lysates were prepared from unirradiated cells & from cells harvested one hour after exposure to 1 or 10-Gy 60Co gamma -radiation & analyzed by immunoblotting with antibodies against several DSB repair proteins. Phospho-ATM & phospho-DNA-PKcs refer to phosphorylation of these proteins at Ser1981 & Ser2056, respectively. Actin served as a loading control. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr2583), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca, Ma

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Ma

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB100-104AF405
Alexa Fluor 488	NB100-104AF488
Alexa Fluor 647	NB100-104AF647
Alexa Fluor 700	NB100-104AF700
Biotin	NB100-104B
DyLight 405	NB100-104V
DyLight 488	NB100-104G
DyLight 550	NB100-104R
DyLight 594	NB100-104DL594
DyLight 650	NB100-104C
FITC	NB100-104F
HRP	NB100-104H
PE	NB100-104PE

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-104SS

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Marker

Modification

Clonality

Clone

Gene Symbol

Images Type

Isotype

Kit Type

Lysate Type

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Protein Or Peptide

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Promotional Item

Species Glossary

53BP1 Antibody - BSA Free

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FANCD2 Antibody - BSA Free

SR-BI Antibody - BSA Free

Nbs1 Antibody - BSA Free

Ki67/MKI67 Antibody - BSA Free

53BP1 Antibody - BSA Free

53BP1 Antibody - BSA Free

CD11b Antibody - BSA Free

ATM Antibody (2C1) - Azide and BSA Free

SR-BI Antibody - BSA Free

ATM Antibody - BSA Free