53BP1: 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.

<b>Left:</b> 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. <b>Right:</b> 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 < 0.0001, MDA-231 to MDA-157, HCC1806, & MDA-468), PAR (**** P < 0.0001, MDA-468 to MDA-157, MDA-231, & HCC1806), & p53 (**** P < 0.0001, MDA-157 to MDA-231, MDA-157 to MDA-468, MDA-157 to HCC1806, MDA-231 to MDA-468, MDA-231 to HCC1806, & MDA-468 to HCC1806). B) Representative fluorescent images used for quantification in A for PARP1, PAR, & p53 for all cell lines tested. Scale bar = 20 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31596905), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: 53BP1 Antibody - BSA Free [NB100-304] - Representative immunohistochemistry staining for 53BP1 expression in pancreatic adenocarcinoma. (A) is high intensity of 53BP1 expression & (B) is low intensity of 53BP1 expression. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23530749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - H2A.J accumulates in the aging human epidermis.(a) H2A.J & Ki-67 co-localization in sections of human skin of the indicated ages by immunofluorescent staining. H2A.J showed mutually exclusive staining with the Ki-67 proliferation marker. (b) H2A.J & 53BP1 foci increase in aging human epidermis. Arrows indicate positively-stained nuclei. A quantification of positive cells for three biological replicates is shown below the immunofluorescence images, & the one-sided Mann–Whitney U-test was used to determine the statistical significance (*P<0.05, **P<0.01). Error bars represent mean±s.e. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms14995), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Depletion of Ajuba results in activation of the DNA damage response. (A) Staining for p53BP1 foci on cells fixed 72 h after transfection with siRNA #3, & siGFP as a negative control. Quantification of the number of p53BP1 foci on three independent siRNA experiments shown in (B). (C, D) Western blots for induction of Chk1 phosphorylation & induction of p53 phosphorylation, showed for the three target sites, 72 h after siRNA transfection. 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.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Depletion of PNUTS & 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 & 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 & red lines mark the mean. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30456372), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - HPV+ HNSCCs have decreased expression of NHEJ & HR proteins including DNA-Pk & BRCA2Cells were treated with mock or 4 Gy radiation, harvested & lysed at 15 minutes post-treatment, & 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. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26336991), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Spontaneous DNA breakage in G1-phase cells: GamGFP shows large spontaneous G1 53BP1 foci to contain multi-break clusters.The large spontaneous 53BP1 foci in undamaged cells, which occur solely in G1 (Harrigan et al., 2011;Lukas et al., 2011a), contain multiple DSBs that are marked by GamGFP. The GamGFP-53BP1 co-localization is more apparent in the absence of Ku. Data are from three (Ku80-proficient) or four (Ku80-defective) independent experiments of >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 < 0.01, ***P < 0.001 using the Kruskal-Wallis test followed by Dunn’s post hoc test. (b) Higher zoom representative images show HB-domain & 53BP1 co-localization, specifically in the euchromatin area. 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] - General chromatin effects of A∆rod & A∆rod∆hbs. (a) DNA staining (DAPI) in U2OS cells expressing A∆rod. Chromatin staining tended to be stronger on or around lamin A∆rod structures. (b) pRb is a known binding partner of lamin A, & so, it was co-stained as a positive control, revealing strong co-localization with the A∆ structures. (c) No co-localization between A∆rod structures & 53BP1 in HeLa cells indicates that chromatin is not damaged by the structures. (d) Similarly, gamma H2AX did not accumulate on A∆ structures. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/2073-4409/6/2/9), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - 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 < 0.01 by Student’s t-test. Source data are provided as a Source Data file Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30886146), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - 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: 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 < 0.0001; n.s. (no significance). a, b Scale bar: 10 µm. c Upon TSQ1 expression, TRF1HP1 alpha results in fewer TIFs compared to Vonly, TRF1 or HP1 alpha controls. TRFHP1 alpha ~11.4% shows decreased TIFs compared to Vonly: ~19.7% ***p = 0.0008; TRF1: ~20.4% ***p = 0.0005; HP1 alpha : ~17.9% *p = 0.0137 (n = 30–38 nuclei). b, c Significance is assessed by one-way ANOVA & Dunnett’s multiple comparison test with 95% confidence level. d Fluorescence images of control cells overexpressing WT hTR (n = 27–36 nuclei). Same color scheme as a. TIFs quantification in the presence of e WT hTR or f Vonly (n = 28–36 nuclei) show minimal baseline DNA damage at telomeres. b, c; e, f Error bars represent s.e.m. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30181605), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Absence of Roma Leads to Activation of the DNA Damage Response & Genomic Instability(A) Representative immunofluorescence analysis of RPA2-pT21, gamma H2AX, & 53BP1 foci in tissue sections from WT & Roma−/− glands at 10dL. The white arrow indicates nucleolar gamma H2AX. Scale bars, 10 μm. The higher magnification panels on the right have been chosen to highlight the foci & are not zoomed images of the left panels.(B) Immunoblot analysis of cell-cycle checkpoint & DNA damage response in extracts from WT & Roma−/− glands at 10dL. Extracts from U2OS cells treated with 5mM HU were used as a damage control.(C) PCR analysis of p21Cip1 & MDM2 & quantitative real-time PCR analysis of p21Cip1 in extracts from 10dL WT & Roma−/− glands (p = 0.0007, Student’s t test).(D) Representative spectral karyotypes (top) & metaphase spreads (bottom panel) prepared from MECs isolated from WT & Roma−/− glands after a full natural wean.See also Figure S2. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27149840), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - A-T post-mitotic neurons are defective in DDR. In a, D30 post-mitotic neurons (D30) were tested by western blot with antibodies specific for the various ATM phosphosubstrates & cleaved PARP at various times after treatment with 5 Gy IR. beta -Actin was used as a loading control. DNA SSBs (b) & DSBs (c) were analyzed in neurons (D30) by alkaline & neutral comet assay, respectively, at two different times after treatment. 20 min after H2O2 or IR treatment was considered the time point with the maximum DNA damage. In b & c, representative photos of comets from untreated cells & at different time points following treatment. The ratio between treated & untreated tail moments of at least 50–70 cells per experimental point is shown in the graphs (values in %). One representative experiment out of three is shown. In d & e, formation & resolution of IR-induced nuclear foci. Neurons (D30) were irradiated with 0.5 Gy, collected at the indicated times & labeled for gamma -H2AX (d) & 53BP1 (e). For each treatment, the number of foci was scored from 100 cell nuclei per duplicate preparations & from three independent experiments (mean±S.D.) (d & e, bottom) For each time point, the difference between Ctrl & A-T was statistically significant (#P<0.01) (analysis performed by the Student's t-test) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25032865), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Exposure to radiation induces persistent genomic instability.(a) Time course of micronucleus frequency in binucleated cells exposed to 1Gy X-rays or Fe ions (average of two independent irradiations, error bars represent SEM). Paired Student's t-test, *p<0.05, **p<0.01. (b) Time course of gamma H2AX- 53BP1 positive foci per cell distribution in a population of 50 cells per condition following exposure to 1Gy of the indicated radiation. One of two experiments is shown. Chi-Square test NI vs. X-rays or Fe ions. day 7 & day 14 p<0.001. Day 21 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 < 0.05; ∗∗∗p < 0.001) were obtained by performing a Student's t test. ND (no cells were detected). Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213671118301528), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - GamGFP marks DSBs in mammalian cells & is inhibited by Ku.(A) GamGFP co-localizes with 53BP1 on laser-induced DNA breaks. (B) Ku inhibits recruitment of GamGFP to laser-induced damage, live cells. (C & D) Ku inhibits recruitment of GamGFP, fixed cells. Mean ± SEM of three experiments, n >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.

$Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Loss of functional PRPF31 induce RNA:DNA hybrid dependent genomic instability but not in mice retinal neurons.(a andb) gamma H2AX & 53BP1 foci analysis in PRPF31 siRNA-transfected RPE-1 cells. (c & d) gamma H2AX & 53BP1 foci analysis in vasculo-stromal fraction derived primary cells fromPrpf31+/A216P mice (Prpf31-ki). (e) gamma H2AX & 53BP1 foci analysis in retina fromPrpf31+/A216P mice on postnatal day 20. All column bars represent the mean. For (a-d) “n”, mentioned on respective column, signify number of cells analyzed from two independent experiments. For (e) n=16 for each column & signify number of retinal sections analyzed; acquired from n=4 eyes. Error bars represent Standard error of Mean (SEM). *P≤0.05; **P<0.01, ***P<0.001 using Mann-Whitney test (a,b), Kruskal-Wallis test followed by Dunn’s post hoc test (c,d); & two tailed unpaired Student’s t-test (e). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30345028), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

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] - Comparative kinetics of 53BP1 & SPOC1 recruitment to radiomimetic drug NCS-induced DSBs. U2OS cells fixed before (mock) or at the indicated times post-treatment (+NCS), immunostained for SPOC1 (green) & 53BP1 (red) & analyzed by confocal microcopy. At the earliest observation period after NCS treatment (15 min) a large number of small 53BP1 stained DSB repair foci emerged, but were not detectably stained by SPOC1. Between 2 to 24 h post-NCS treatment the number of small 53BP1 stained foci gradually, then almost completely disappeared, concomitant with emerging larger repair foci. At around 8 h post-NCS treatment most of the large 53BP1-stained repair foci were also strongly stained by SPOC1, suggesting SPOC1 is recruited to & accumulates at DSBs in heterochromatin known for their slow repair kinetics. Scale bars = 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.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - RNA:DNA hybrid formation is central to ATM-53BP1 repair pathway.(a) Immunofluorescence using the anti-flag to quantify nuclear hybrid-binding (HB) domain foci signal in pre-permeabilized RPE cells & quantification of HB domain signal in cells treated with siATM, (b) Western blot showing ATM depletion. (c) Immunofluorescence & HB-domain foci quantification in cells treated with ATM inhibitor. (d) Quantitation of gamma H2AX foci in cells expressing HB domain or active RNAseH1, in presence or absence of H2O2 dependent oxidative stress. siC, non-targeted control siLuciferease RNA. 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 < 0.01, ***P < 0.001 using Mann-Whitney test (a,c) & Kruskal-Wallis test followed by Dunn’s post hoc test (d). 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] - UMI acidic residues are required for RNF168-mediated site-specific ubiquitination & the DDR.a Sequence alignment analysis of RNF168 UMI domains across species. The evolutionary conserved E143/E144 are highlighted in red. b Deletion of residues 143-144 attenuates 53BP1 IRIF formation. U2OS RNF168 KO cells were transiently transfected with GFP-RNF168 or RNF168 delta 143-144. Cells were irradiated with 10 Gy, allowed to recover for 1 h, followed by immunofluorescence analysis with indicated antibodies. Repeated three times with similar results. c Deletion of 143-144 abolishes RNF168-mediated ubiquitination. Co-transfection of GFP-RNF168 or mutant with Flag-H2AX K13/K15 only vector in HEK293T cells followed by western blot analysis. Repeated three times with similar results. d RNF168 delta 143-144 is recruited to laser damage. U2OS RNF168 KO cells expressing GFP-RNF168 & RNF168 delta 143-144 were damaged using a 405 nm laser source & analyzed 15 min later by confocal microscopy. Dotted line indicates the laser path. Repeated three times independently with similar results. e–f U2OS cells were transfected with GFP-RNF168 wildtype & mutants. Proximity ligation assay (PLA) was performed using specific antibodies against GFP & gamma H2AX. Mean intensity of PLA signals in GFP-positive cells were measured & analyzed using ImageJ software. Data presented as mean ± SD. One-way ANOVA, Dunn’s multiple comparison test was used in statistical analysis, wildtype vs. R57D P = 0.1455, wildtype vs. delta 143-144 P = 0.1589, R57D vs. delta 143-144, P > 0.9999, ****P < 0.0001 with 95% confidence interval. n = 75 (wildtype), 48 (R57D), 74 ( delta 143-144), 53 (R57D/ delta 143-144) over 3 independent experiments. Source data are provided as Source Data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32424115), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - TRF2/Terc double null mutant mice exhibit DNA damage response at short telomeres in epidermisA. Average telomere length ratios in CD34+ stem, Lgr6+ stem, & basal cells from K14Cre;TRF2+/+;Terc+/+, K14Cre;TRF2f/f;Terc+/+, K14Cre;TRF2+/+;Terc-/-, & K14Cre;TRF2f/f;Terc-/- epidermis were determined by qPCR. Error bars represent SEM. 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+/+ B., K14Cre;TRF2f/f;Terc-/- C., K14Cre;TRF2+/+;Terc-/- D., & K14Cre;TRF2f/f;Terc+/+ E. epidermis is shown. Nuclei are counterstained with DAPI. Scale bar = 5 μm. Phospho-ATM expression in histopathologic sections from K14Cre;TRF2+/+;Terc+/+ F., K14Cre;TRF2f/f;Terc-/- G., K14Cre;TRF2+/+;Terc-/- H., & K14Cre;TRF2f/f;Terc+/+ I. epidermis. Phospho-Chk2 expression in histopathologic sections from K14Cre;TRF2+/+;Terc+/+ J., K14Cre;TRF2f/f;Terc-/- K., K14Cre;TRF2+/+;Terc-/- L., & K14Cre;TRF2f/f;Terc+/+ M. epidermis. p53 protein expression in histopathologic sections from K14Cre;TRF2+/+;Terc+/+ N., K14Cre;TRF2f/f;Terc-/- O., K14Cre;TRF2+/+;Terc-/- P., & K14Cre;TRF2f/f;Terc+/+ Q. epidermis. Representative sections 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.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Vismodegib increases radiation-induced gamma H2AX/53BP1 nuclear foci. BCC-1 or SCC-25 cells were treated for 24 h with 10 or 40 µM vismodegib or with DMSO as control followed by irradiation with 4 Gy. (A) Representative images of phosphorylated gamma histone-2AX ( gamma H2AX)/p53-binding protein 1 (53BP1) foci from non-irradiated & 4 Gy-irradiated DMSO control versus 40 µM vismodegib-treated cells are shown. Nuclei were counterstained with DAPI. Scale bar, 10 µm. (B) Quantification of persistent gamma H2AX/53BP1 foci in vismodegib treated & irradiated BCC-1 (left panel) & SCC-25 (right panel) cells. Bars represent means + SD from three independent experiments performed in duplicate. ** p < 0.01 vismodegib- versus DMSO-treated cells & #p < 0.05, ##p < 0.01 4 Gy versus non-irradiated cells (t-test). Gy, Gray; Vism., vismodegib. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30142876), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - The hypoxia-activated pro-drug Tirapazamine (TPZ) reduces stem-like Aldefluorbright population in vitro & in liver metastases(a) Human colonospheres cultured in hypoxia (0.1%) & normoxia (21%) for 24 hrs in absence or presence of TPZ for 4 hrs. Cells processed for IF analysis of gamma H2AX (green). DAPI (blue) used to visualize cell nuclei. (b) Bar graphs showing quantification of gamma H2AX foci observed in (a), shown as % of mean gamma H2AX intensity at normoxia (n=3). (c) Representative confocal pictures of comet assay of colonospheres cultured as in (a). (d) Bar graphs showing quantification of DNA damage observed w/ the comet assay (Extend tail moment) (n=77). (e) Experiment performed as in (a), but cells either treated or untreated for 16 hrs. Cells lysed & analyzed by WB for the indicated markers (cropped). (f) Colonospheres cultured in hypoxia & treated w/ TPZ for 16 hrs. The cells FACS sorted into Aldefluorbright & Aldefluordim populations & analyzed by WB for the indicated markers. (g) Human colonospheres cultured in hypoxia (0.1%) for 24 hrs in absence or presence of TPZ for 4 hrs & FACS sorted into Aldefluorbright & Aldefluordim populations. Bar graph the fold change Aldefluordim & Aldefluorbright cells upon TPZ treatment (n=3). (h) C26 tumor cells injected into liver parenchyma of immune-deficient mice. Following tumor initiation, tumor-bearing liver lobes subjected to a vascular clamping protocol [24] to induce hypoxia & treated w/ saline (control) or TPZ (treatment) for 10 days. At endpoint, livers excised & expression of ALDH1 examined by IHC. Magnification 20x, T=tumor. (i) Bar graphs showing quantification of ALDH1 (n=28) observed in (h) (n=14). * = significant (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.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - MRE11 poly(T)11/12 sequence variant has no apparent functional consequence on DNA damage detection & repair.(A) Electropherograms & sequencing traces of the MRE11 poly(T)11 tract in leukemia/lymphoma cells: Daudi (heterozygous insertion of one bp, poly(T)11/12); Jurkat (heterozygous deletion of one bp, poly(T)10/11); & Raji (wild-type, poly(T)11). (B) MRE11 expression in Daudi, Jurkat & Raji cells. qRT-PCR was used to calculate the ratio of MRE11 expression (amplification of exons 2 to 4) relative to the GAPDH control, independently of the presence of the aberrant transcript observed in Jurkat. PCR products were loaded on the gel at saturation of the reaction, being not quantitative. MRE11 protein levels were assessed by Western blot, & were considerably lower in Jurkat. (C) ATM-pS1981 levels were assessed by Western blot in Daudi, Jurkat & Raji after 1.5 Gy irradiation of cells & 1 h, 4 h or 24 h recovery. There was an increase of ATM phosphorylation 1 h after irradiation in Daudi & Raji cells that decreased at the later time points. Jurkat showed no ATM phosphorylation after exposure to irradiation. (D) DNA repair foci was observed by immunofluorescence after 1.5 Gy IR followed by 1 h & 4 h recovery. Irradiation resulted in the formation of both MRE11 & 53BP1 foci in Daudi & Raji cells, whereas Jurkat failed to form MRE11 foci. Nuclei were counterstained with DAPI. Merged figure presents co-localization of MRE11 & 53BP1 foci visualized in yellow. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21637783), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Cry2−/− cells accumulate damaged DNA.(A) Representative early passage (P3–4) primary wildtype (WT), Cry1−/−, Cry2−/−, & Cry1−/−;Cry2−/− adult ear fibroblasts stained with anti-53BP1 antibody (green) & DAPI (blue). Insets show enlarged view of indicated cells. (B) Quantitation of 53BP1-positive cells prepared as described in (A). Nuclei containing more than five 53BP1 puncta & negative for BrdU labeling were considered positive for DNA damage. Data represent the mean ± s.d. for at least 200 cells per genotype. (C) Chi-squared analysis of the distributions of Cry1 & Cry2 wildtype (black), heterozygous (gray), & homozygous knockout (red) genotypes establishes a significantly reduced survival of Cry2−/− mice. **p < 0.01 by chi-squared analysis with 2 degrees of freedom (χ2 = 10.39).DOI://dx.doi.org/10.7554/eLife.04883.020 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25756610), 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 < 0.05, ****P < 0.00005. (B) Immunoblots for the detection of gamma H2AFX & histone H4. Right panel shows gamma H2AFX/H4 ratios at indicated pHe values. C, control before damage induction. Quantitation of three independent immunoblots is shown on the right. Values are mean ± SEM. *P < 0.05. (C) Immunofluorescence detection of gamma H2AFX after 48‐h incubation in range of pHe values tested without BLM treatment. Right panel corresponding to gamma H2AFX foci/cell at 48 h of pHe incubation. Only foci larger than 10 pixels were counted. Experiments were performed in triplicate. Values are mean ± SEM. *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29511618), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - BETd blocks 53BP1 function in senescent cells.a Early passage (control) or late passage (replicative senescence) TIG-3 cells were treated with 10 nM ARV825 (+) or vehicle (−) for 4 days. These cells were then subjected to western blotting using the antibodies shown on the right after immunoprecipitation with the antibodies shown at the top of the panel (IP:). b–f Early passage (control) or late passage (replicative senescence) TIG-3 cells were transfected with previously validated two different siRNA oligos against 53BP1 or control siRNA oligo twice at 2 days intervals. These cells were then subjected to RT-qPCR analysis (b), western blotting analysis (c), the cell proliferation analysis (d), immunofluorescence staining using the antibodies shown on the left (e) or neutral comet assay (f). Representative photographs of the cells in the indicated culture conditions are shown & the histogram shown at the bottom of the panel indicates the relative cell number (d). The number of gamma H2AX foci, above threshold intensity per nucleus (n = 50) was quantified & shown at the bottom of the e. The average tail olive moments were shown at the bottom of the f. Each box indicates the median & 25% & 75% quantiles, & the whiskers represent the minimum & maximum observations (e, f). For all graphs, error bars indicate mean ± s.d. (n = 3 for b, n = 4 for d) & the representative data from three independent experiments were shown. Statistical significance was determined with two-tailed unpaired Student’s t-test (b), one-way ANOVA followed by Tukey multiple comparison test (d), one-way ANOVA followed by Holm-Sidak multiple comparison test (e, f). P values < 0.05 were considered significant. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32321921), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Bystander signaling evokes DNA damage foci formation in urothelial cancer & normal urothelial cells(A) Schematic representation of experimental conditions including non-irradiated, uniformly-irradiated & shielding protocols where 50% of the slide was exposed & 50% shielded with MCP alloy. Cells were fixed & processed for 53BP1-immunofluorescence, one hour following irradiation. Micrographs of urothelial cancer T24 cells labelled with anti-53BP1 (green) to label nuclear DNA double stand break foci & DAPI to counterstain nuclei are shown for each experimental condition. (B) Summary graph of mean foci/cell in T24 cells (N=3), measured from confocal z-stacks in Volocity software. There were significantly more foci in shielded cells 0.5mm from the shield than in non-irradiated controls, indicative of a bystander effect (p=0.019). (C) Summary data for HT1376 cancer cells (N=3) where there was no difference in the mean foci/cell between non-irradiated controls & shielded cells. (D) Summary data for normal urothelial HUC (N=3). Mean foci/cell was significantly greater in shielded cells in the 0-5mm (p=0.005) & 5-10mm (p=0.04) analysis regions from the shield. * denotes p<0.05, ** denotes p<0.01 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29228614), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Pot1a prevents DDR in HSCs. a, b Telomeric DDR in 8 week-old LSK cells upon Pot1a knockdown. a Immunocytochemical staining of TRF1 (green) & 53BP1 (red). Foci co-stained with TRF1 & 53BP1 were identified as TIFs. Nuclei were stained with TOTO3 (blue). Scale bar, 2 μm. b Frequency of TIFs per cell after 1 week of culture. Data are expressed as the mean ± SD (n = 100, *p < 0.01 by t-test). Representative data from three independent experiments are shown. c–f Telomeric DDR in donor-derived control-GFP or Pot1a overexpressing 8 week-old LSK cells 4 months post BMT. c Immunocytochemical staining of TRF1 (green) & 53BP1 (red). Nuclei were stained with TOTO3 (blue). Scale bar, 2 μm. d Frequency of TIFs per cell. Data are expressed as the mean ± SD (n = 120, *p < 0.01 by t-test). Representative data from 3 independent experiments are shown. e Immunocytochemical staining of TRF1 (green) & RPA32 (red). f Immunocytochemical staining of TRF1 (green) & pChk1 (red). Scale bar, 2 μm. g Flow cytometric analysis of Chk1 & pChk1 in donor-derived GFP+ LSKCD48–CD150+ cells after 5 months of 2nd BMT. Mean fluorescence intensity of Chk1 & pChk1 (left panels). Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative FACS profiles of Chk1 & pChk1 in donor-derived GFP+ LSKCD48–CD150+ cells (right panels) 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.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Aphidicolin-induced ATM signaling in immortalized MEFs. (A) Cells (WT, MJI-53; KO, MJI-1) were treated for 24 h using 0 or 3 μM aphidicolin. (B) Cells (lane 1–3, WT, MJI-91; lanes 4–6, KO, MJI-186; lanes 7–9, WT, MJI-105; lanes 1–12, KO, MJI-84) were treated for 24 h using 0, 1 or 3 μM aphidicolin. Actin & 53BP1 serve as loading controls. DYNLL1 is used as a surrogate marker for loss of ASCIZ. (C) Quantification of western blot band intensities. An arbitrary unit of 100 represents the average band intensity for the respective phospho-protein in the 3 μM aphidicolin-treated wildtype samples on each membrane. Graphs indicate the mean ± standard error, n = 3. Additional loading controls for total KAP1, p53 & H2AX are shown in Supplementary Fig. S1A & B. 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] - POT1 prevents DDR in human HSC. a Expression of POT1 in hCB CD34+ cells & LT-HSCs (Lin–CD34+CD38−CD45RA−CD90+CD49f+). b hCB-derived LT-HSCs were cultured with MTM-POT1. After 10 days of culture, cells were isolated & re-cultured in methylcellulose medium (200 cells per dish). The number of CFU-C & HPP-CFC (>1.0 mm, >2.0 mm) are shown. Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative data from three independent experiments are shown. c, d hCB LT-HSCs were cultured for 10 days with control MTM protein or MTM-POT1. After 10 days of culture, LT-HSCs were re-isolated & number of TIF was examined. c Immunocytochemical staining of TRF1 (green), 53BP1 (red), & TOTO3 (blue). Scale bar, 2 μm (left). Frequencies of TIFs after 10 days of culture (right). Data are expressed as the mean ± SD (n = 100: control, n = 100: MTM-POT1, *p < 0.01 by t-test). Representative data from 2 independent experiments are shown. d Immunocytochemical staining of TRF1 (green), RPA32 (red), & TOTO3 (blue). Scale bar, 2 μm (left). Frequencies of TIFs after 10 days of culture (right). Data are expressed as the mean ± SD (n = 110–120: control, n = 110: MTM-POT1, *p < 0.01 by t-test). Representative data from two 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] - Telomere-initiated senescent cells retain active DDR foci for years after senescence establishment.a. DDR, in the form of ATM pS1981 foci co-localizing with 53BP1 & gamma H2AX foci, is detectable three years after senescence establishment. Scale bar, 10 µm. Below, bar graphs show the percentage of cells positive ± s.e.m. for the indicated DDR markers, in senescent (sen), early passage proliferating (prol) or telomerized proliferating (tel) skin fibroblasts from two independent centenarian donors (cen2 & cen3). Cells were considered positive if bearing more than 3 DDR foci (*** p-value <0.001). b. SA-beta -gal staining of the two batches, cen2 & cen3, is shown together with the percentage of BrdU-positive cells. Scale bar, 100 µm. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0110969), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - VPA at a safe dose can lead to the dysfunction of DNA repair function. (A) The images represent gamma H2AX & 53BP1 foci formation in U2OS cells treated with 0.5 or 1.0 mM VPA at 24 h post-IR treatment (left panel), & the percentage of cells with gamma H2AX foci or 53BP1 foci formation in each group was calculated (right panel, the cell with >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 < 0.05). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28489060), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - The susceptibility of human LECs to low-dose ionizing radiation. The human lens cell line FHL124 was exposed to low-dose IR up to 2.28 Gy. Exposed cells were then processed for both immunofluoresence microscopy (a) & immunoblotting (b) 1 h later. The signals obtained by immunoblotting were quantified & the mean from three independent experiments calculated & plotted (c) against IR dose. GAPDH was used as a loading control. Both gamma H2AX & RAD51 increased linearly with IR dose. Signals for other markers of DNA repair, 53BP1, MRE11 & TP53, remained unchanged as assessed by immunoblotting (b). By immunofluoresence microscopy (a), MRE11 & 53BP1 redistributed into nuclear foci, particularly at the 2.28 Gy level. TP53 remained uniformly distributed throughout the nuclear compartment, but excluded from nucleoli (a). As the levels of gamma H2AX & RAD51 increased after exposure to IR (c), so the number of nuclear foci also increased (a). Scale bar, 10 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25924630), 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] - UDM1 is required for RNF168-mediated 53BP1 recruitment.a Schematic illustration of RNF8 & RNF168 structural domain organization, RNF168 fragments, & chimeric proteins used as in b & c. FHA–forkhead-association domain; coiled-coil–coiled-coil domain; RING–ubiquitin E3 ligase RING domain; Rs-arginine anchor; LRM–LR motif; UMI–UIM-and MIU-related ubiquitin binding domain; MIU–motif interacting with ubiquitin. b–c Transient transfection of GFP-tagged RNF168 fragments & chimeric proteins in U2OS RNF168 KO cells. Cells were irradiated with 10 Gy & allowed to recover for 1 h followed by immunofluorescence analysis using 53BP1 antibody as indicated. Cells were counterstained with DAPI. Repeated at least two times independently with similar results. Source data are provided as Source Data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32424115), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Loss of ATM function in retina.(a) Retinal explants pretreated with ATM inhibitor (ATMi) or PI3K inhibitor (DDRi) are irradiated & sections analyzed for gamma H2AX. (b) ATM knockout adult mice retina analyzed for irradiation dependent gamma H2AXaccumulation. (c) Western blot analysis of ATM & 53BP1 in micro-dissected neuronal retina. LC, Coomassie-equivalent staining of gels used as loading control (representation of n=2 independent repeats). (d) Cartoon depiction of domains of ATM protein. (e) Neural retina of mice was microdissected & cDNA was prepared. PCR performed using indicated domain-specific primers onATM mRNA. Unlike FATC & TAN domain coding mRNA; the FAT-domain containing mRNA could be observed even in 5-month-old adult mice retina, indicating the presence of an alternatively spliced form of ATM in the retina. 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] - TIFs measured in leukocytes. (A) The percentage of cells with one or more TIFs in griffon vultures. The age & number of cells measured for each individual was as follows: 8.06 years: 77 cells, 10 years: 29 cells, 10.9 years: 153 cells, 12.3 years: 146 cells, 15.3 years: 125 cells, & 21.4 years: 173 cells. (B) The percentage of cells with one or more TIFs in American flamingos. The age & number of cells measured for each individual was as follows: 0.79 years: 3 cells, 1.75 years: 12 cells, 1.8 years: 34 cells, 6.75 years: 21 cells, 13.8 years: 11 cells, 16.9 years: 9 cells, 19.9 years: 7 cells, 19.9 years: 7 cells, 34.9 years: 20 cells, & 35.9 years: 6 cells. In the representative images, the nuclei are stained blue with DAPI, the telomeres are red, & the 53BP1 stain is in green. 53BP1 staining at the very edge of nuclei was not counted as foci. The top row of the representative images shows maximum projections which are the result of taking the maximum value of several different z-planes. The magnification image is displayed for a single plane rather than a maximum projection to show co-localization of the stains. A white arrow indicates a colocalization of 53BP1 & a telomere spot (a TIF). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31730540), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - NRK1 deletion curbs PARP activity & exacerbates DNA damage. a NAD+ levels in the liver of NRK1 LKO mice upon LFD & HFD (n = 8 for all groups except NRK1 LKO on HFD, n = 10; 24 weeks of age). b NR (left), NMN (center) & NAM (right) levels in the liver of NRK1 LKO & control mice upon HFD (n = 6 mice per group). c mRNA level of genes involved in the NAD+ biosynthesis pathways (n = 8 mice for the control group, n = 10 mice for the NRK1 LKO group). d Levels of poly(ADP-ribose) protein modification, PARP1, SIRT1, NAMPT, & GAPDH in the liver of NRK1 LKO & control mice on HFD. e Representative staining (top) & quantification (bottom) of gamma H2AX (left) & 53BP1 (right) immunofluorescence on liver sections from NRK1 LKO & control mice on HFD (n = 4 mice per group, ×10 magnification, DAPI counterstaining, scale bars: 200 μm). Results shown are mean ± SEM, * & *** indicate statistical difference vs. control group at p < 0.05 & p < 0.001, respectively. The individual values & statistical tests used for each panel can be found in the Source Data file Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31541116), 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, Po, Pm, Fe, Pm-Or, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, 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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NB100-305

53BP1 Antibody - BSA Free

(6 Reviews)

(152 Publications)

0.1 mg

NB100-305

<b>Biological Strategies Validation. </b>53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-305-img0008.jpg

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.

<b>Biological Strategies Validation. </b>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. Dotted line represents young level. BER: Base Excision Repair, NER: Nucleotide-Excision Repair; MR: Mismatch Repair, DRR: DNA Repair Related. (D–E) Satellite cells freshly isolated from uninjured (D) or 72 hrs after injury (E) muscle of C57BL/6 young & old mice, & of young SCID mice. Cells plated at low density, irradiated at indicated Gray (Gy) doses, & cultured for 10 days. Myogenic colonies formed by irradiated cells quantified & represented relative to ir respective non-irradiated controls. On average, 178, 77, & 228 myogenic colonies scored per mouse for young, old, & SCID uninjured non-irradiated respectively & 67, 34, & 91 myogenic colonies scored per mouse for young, old, & SCID injured non-irradiated respectively. Data represent mean +/− SEM, n = 3 to 4 mice per group, two-tailed unpaired Student's t-test, a, b or c: p value<0.05, (a: young vs old, b: young vs SCID, c: old versus SCID). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23704914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

PARP1-dependent end joining repairs the resected DSBs in 53BP1- or RIF1-depleted G1 cellsA. HeLa cells harboring pEJ were treated with the indicated siRNAs for 24h and PARP1 was then inhibited by adding 1μM olaparib 2h before transfection with I-SceI-expressing vector. The percentages of GFP+ cells were measured after 48h as indication for end joining efficiency. PARP1-EJ was measured as the percent of suppressed end joining after PARP1 inhibition. B. Representative micrographs for gamma H2AX foci at 8h post 2Gy in A549 cells after depletion of either 53BP1 or RIF1 proteins in the presence or absence of olaparib. Nuclei were counterstained with CenpF to determine G1 (CenpF−) cells. C. Quantitation of gamma H2AX foci number at the indicated time points in CenpF− G1 cells. D. Left panel: Western blot showing an efficient siRNA-mediated knockdown of the indicated individual or combined proteins. Right panel: Asynchronous A549 cells were treated with colcemid (0.2μg/ml) for 1h pre-2Gy and the number of gamma H2AX foci was monitored in CenpF− G1 cells after the indicated treatment conditions. At least 50 nuclei were counted. (**) indicates P < 0.001. In all cases, the number of foci measured in non-irradiated cells was subtracted (relative). Shown are the mean +/-SEM for three independent experiments. CTR: control, Olap: olaparib, sc: scrambled RNA, si53: si53BP1, and siR: siRIF1. 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.

CtIP and MRE11 but not EXO1 are required for end resection in 53BP1-depleted G1 cellsA. A549 cells were treated as in Figure 4 with the indicated siRNAs and efficient knockdown of 53BP1 and/or EXO1 was confirmed by Western blot (upper panel). Lower panel: The focus intensity of RPA was measured in 53BP1- and/or EXO1- depleted cells at 2h post-2Gy in G1 and S/G2. B. Immunoblots showing efficient siRNA-mediated knockdown of CtIP and/or 53BP1 (upper panel), MRE11 and/or 53BP1 (middle panel), or PLK3 and/or 53BP1 (lower panel). (C & D) Quantitation of the percentage of G1 cells with more than three C. RPA or D. CtIP foci after the indicated treatments. At least 50 nuclei were counted. (**) indicates P < 0.001, and (ns) means P > 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 < 0.001. (D and E) Serum-starved WI38 hTERT cells were treated with DMSO or MG132 (50 μM) for 1 h before exposure to 0.8 Gy IR. One hour post-irradiation, cells were co-stained for gamma H2AX (green) and 53BP1 (red). (D) Representative pictures. (E) Number of gamma H2AX foci per nucleus from one representative experiment (162–180 nuclei were analyzed for each treatment) out of three. Ns: not significant. (F and G) U2OS EV28 cells were treated with DMSO or MG132 (10 μM) for 1 h before the addition of ethanol (untreated) or 300 nM 4-hydroxitamoxifen (4OHT) for 4 h to express AsiSI in the nucleus (42). (F) Representative pictures of cells co-stained for gamma H2AX (green) and 53BP1 (red). (G) ChIP analysis using an anti-gamma H2AX antibody (black) or a non-immune antibody (IgG, gray). Enrichment was assessed by QPCR amplification using primers proximal to two AsiSI sites located inside two genes (Gene I: SFRS6, Gene II: CCD47) and primers distal to an AsiSI site (Control). Enrichment was normalized to the maximum recovery for each experiment (means +/- SEM, n = 3). Ns: not significant; *P < 0.05. In the microscopic images, nuclear contours, identified by DAPI staining (not shown), are indicated by dashed lines. Bars: 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26578593), 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.

Exosomes modulate the repair of DNA DSBs in irradiated recipient cells.(A) Number of 53BP1 foci in BHY cells 1 hour after irradiation with 0 and 2 Gy and transfer of BHY exosomes isolated 24 hours after irradiation with 0 and 6 Gy [n = 5]. (B) 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). (C) Number of 53BP1 foci in BHY cells 6 hours after 2 Gy and transfer of BHY exosomes isolated 24 and 48 hours after irradiation [n1 (control; EXO 0 Gy 24 h; EXO 6 Gy 24 h) = 6, n2 (EXO 0 Gy 48 h; EXO 3 Gy; EXO 6 Gy 48 h; EXO 9 Gy) = 3]. (D) Number of 53BP1 foci in FaDu cells 6 hours after 2 Gy and transfer of FaDu exosomes [n = 3]. (E) Number of 53BP1 foci in FaDu cells 6 hours after 2 Gy and transfer of BHY exosomes [n = 3]. (F) Number of 53BP1 in BHY cells after 2 Gy and transfer of destabilized BHY exosomes. Exosomes from BHY cells isolated 24 hours after irradiation with 0 and 6 Gy were treated with RNase A or a mixture of Triton and Trypsin [n1 (control; intact) = 6; n2 (RNase A 5 μg/μl) = 2; n3 (RNase A 400 μg/μl; Triton + Trypsin) = 3]. For all experiments the +/- SD was shown and p-values calculated on control were considered to be significant if * p < 0.05 and highly significant ** if p < 0.01, while ▲ p < 0.05 and ▲▲ p < 0.01 indicate significant differences to EXO 0 Gy. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27006994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Inhibition of ACLY exacerbates radiation induced DNA damage in HNSCC cells and impairs homologous recombination. (A,B) UM-SCC-6 (A) and UT-SCC-5 (B) cells were treated with 5 uM BMS303141 for 6 h and irradiated with 2, 4, or 6 Gy. Survival was determined by colony formation assay. (C,D) Quantification of cells positive for 53BP1 foci (>5 53BP1 foci/nucleus in a single plane of view) in UM-SCC-6 (C) and UT-SCC-5 (D) cells. Shown are mean percentages of three biological replicates. Immunofluorescent representative images of examples of UM-SCC-6 or UT-SCC-5 cells, 24 h post irradiation, with or without 5 uM BMS303141 pre-treatment. Nuclei in blue, 53BP1 foci in red. (E) Homologous recombination assay. U2OS cells have been stably transfected with a construct containing a GFP gene containing a I-SceI restriction site (SceGFP) as well as a stop codon at the same site. Upon transfection with an I-SceI encoded plasmid, a double strand break is induced at the I-SceI site. Repair of this break via homologous recombination occurs through utilization of the internal GFP (iGFP) fragment downstream of the SceGFP. Successful homologous recombination results in the restoration of a functional GFP gene. (F) U2OS DRGFP cells were treated for 6 h with 5 uM BMS303141 or 5 uM BO2 (Rad51 inhibitor), and transfected with pCBASceI plasmid. 48 h post transfection, cells were harvested and the GFP positive fraction was determined using flow cytometry. Representative images from flow cytometry experiments are shown on the right, with DMSO Sce- as a negative control, and 5 uM B02 Sce+ as a positive control. In the graph, data are shown from two biological replicates. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31817870), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

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 < 0.05, **P < 0.01). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.5423), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Cellular RER & not enzyme activity against RNA:DNA hybrids correlates with DNA damage & proinflammatory responseAOverexpression of RNase H1 in Rnaseh2b−/− cells restores RNase H activity against RNA:DNA hybrids to 81 ± 10% of wild‐type levels, while overexpression of RNASEH2B restores cellular enzyme activity for cleavage of both RNA:DNA & DRD:DNA substrates (RER). Rnaseh2b−/− MEFs were complemented with Rnaseh1 (+H1), Rnaseh2b (+H2B) or EGFP by retroviral infection. Mean of n = 3 independent experiments ± SEM.B, CDNA damage is reduced to wild‐type levels by complementation with Rnaseh2b but not Rnaseh1, measured by 53BP1 foci formation in detergent‐extracted fixed cells. (B) Representative images (scale bar, 10 μm). (C) At least 150 cells were counted for each cell line in three independent experiments. Mean ± SEM, ****P < 0.0001 two‐tailed t‐test.D–FCCL5 (D) & CXCL10 production (E), as well as ISG induction (F) in Rnaseh2b−/− MEFs are reduced close to wild‐type levels (Rnaseh2b+/+), by complementation with Rnaseh2b but not Rnaseh1. Mean of n = 6 independent experiments ± SEM for complemented cells; n = 3 independent experiments for Rnaseh2b−/− parental & Rnaseh2b+/+ controls cells. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 two‐tailed t‐test indicates significantly reduced expression compared to Rnaseh2b−/− parental cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26903602), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Correlation of CK2 & PTEN subcellular distribution with delay in DNA damage response in GBMs.(a) Co-IF of CK2 beta & PTEN with DAPI nuclear staining in the absence of doxorubicin (DOX−; dimethylsulfoxide (DMSO)-treated) or after 5 h of exposure to DOX (DOX+; 0.5 μM DOX-treated) in GXP GBM3 compared with WT or GXP NS. Representative images are shown. See Supplementary Fig. 3a for representative IFs of GX or GP NS, GXP GBM2 & T98G. Scale bar, 20 μm. (b) Immunoblot analysis of nuclear (N) & cytosolic (Cy) distributions of total PTEN & S380/T382/T383-phospho-PTEN in WT or GXP NS, or GXP GBM3 in DOX− (DMSO-treated) & DOX+ (0.5 μM/5 h) conditions. Loading controls: S6RP (cytosolic) & Histone3 (nuclear). (c) Immunoblot analysis of PTEN & CK2 beta in two representative GXP GBMs compared with GXP NS in DOX− (DMSO-treated) & DOX+ (0.5 μM/5 h). Loading control: beta -actin. (d) Co-IF staining of DOX-induced foci of gamma H2AX & 53BP1 in WT or GXP NS, GXP GBM2 & T98G. Cells were exposed to DOX (0.5 μM) for 0, 0.5, 1 or 5 h, & allowed to recover for 5 or 24 h before fixation, & antibody & DAPI nuclear staining. Representative images are shown. Scale bars, 10 μm. IF images for GX or GP NS compared with WT NS are further shown in Supplementary Fig. 3b. The percentage of gamma H2AX-positive cells (y axis) at the indicated time points (x axis) for each group is summarized in the graph in (e). Error bars represent mean±s.d. Representative 100 cells were randomly selected for quantification. Cells containing ≥5 foci were considered as gamma H2AX positive. See also Supplementary Fig. 3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28094268), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Recruitment of DNA repair factors to double-strand breaks is impaired in splicing-deficient cells. (a) HeLa Luc-I or Luc cells were exposed to DMSO (control), 100 nM pladienolide B or 50 μM isoginkgetin for 1–16 h. After normalization to the corresponding DMSO value, the ratio between Luc-I & Luc luciferase activity is presented as a percentage. Means±S.D. are shown, n=4. (b) U2OS cells were treated with pladienolide B or isoginkgetin for 2, 6 or 16 h, irradiated (6 Gy, 1 h recovery) 1 h prior to termination of the treatment, fixed & immunostained for gamma H2AX, MDC1, WRAP53 beta , RNF168, conjugated ubiquitin recognized by the FK2 antibody, 53BP1, RAD51 or BRCA1. Nuclei were stained with DAPI in all immunofluorescence experiments. The numbers in white represent the percentage of 100–200 cells counted whose nuclei contained >10 IR-induced foci. Means±S.D. are shown, n=3. *P-value<0.05, as determined by a non-paired two-tailed Student's t-test. The ‘foci-like' accumulations RAD51 after splicing inhibition in (b) are not IR-induced foci, but accumulation of RAD51 in the nucleolus for unknown reasons Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27315300), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-904] - Inhibition of splicing downregulates repair factors at both the mRNA & protein levels. (a) U2OS cells were treated with pladienolide B for 2, 6 & 16 h & irradiated (6 Gy, 1 h recovery) 1 h prior to termination of the treatment. The levels of the indicated mRNAs were measured through qPCR analysis. The change is relative to the DMSO control & two reference genes (18S rRNA & beta -actin). Means±S.D. are shown, n=3. (b) qPCR analysis of the levels mRNA & pre-mRNA for RNF8 & RAD51 in U2OS cells treated with pladienolide B or isoginkgetin for 2, 6 or 16 h & irradiated (6 Gy, 1 h recovery) 1 h prior to termination of this treatment. The change is expressed relative to the DMSO control value & the levels of mRNA for two reference genes (18S rRNA & beta -actin). Means±S.D. are shown, n=3. The arrows indicate the positioning of the PCR primers used & their sequences are shown in Supplementary Table S1. (c) Western blotting following pladienolide B treatment of U2OS cells as described in (a). beta -Actin was used as a loading control. Densitometric quantification of each protein is shown in Supplementary Figure S2B. The three MDC1 bands correspond to the unphosphorylated, phosphorylated & hyperphosphorylated forms of full-length MDC1 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27315300), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: 53BP1 Antibody - BSA Free [NB100-904] - Reptin depletion reduces the recruitment on chromatin of BRCA1 & 53BP1 after gamma ray irradiation.HuH7 cells stably expressing a doxycycline-inducible Reptin shRNA were treated with doxycycline (sh Reptin +) or left untreated (sh Reptin-) for 4 days. (A) Representative images of 53BP1 & BRCA1 foci in HuH7 cells detected using immunofluorescence 2h after gamma ray irradiation. (B) The bars represent the mean number of foci per cell from two independent experiments (>200 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. *P<0.05, as determined by a non-paired two-tailed Student's t-test Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27315300), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Restoration of nuclear PTEN-mediated DNA damage signalling by restraining CK2 impairs tumour but not normal neural stem cell survival.(a) Immunoblot analysis displaying the impact of TBB treatment (25 μM/24 h) on CK2 beta , PTEN, p-PTEN (S380/T382/T383), p-AKT (S473) & p-PRAS40 (T246) in GXP GBM3 & 4, with beta -actin used as loading control. (b) IF of PTEN with DAPI nuclear staining in dimethylsulfoxide (DMSO)- or TBB (25 μM/24 h)-treated cells in the absence of DOX or after 5 h of exposure to DOX (0.5 μM). DOX was added 5 h before the end point of DMSO or TBB treatment. Representative images are shown. Scale bars, 20 μm. (c) Co-IF of gamma H2AX & 53BP1 foci formation with DAPI nuclear staining in DMSO- or TBB (25 μM/24 h)-treated GXP NS or GBM2 with DOX−, or 1 h or 5 h of DOX (0.5 μM) exposure followed by 5 or 24 h of recovery time. Scale bars, 10 μm. A graph quantifying the percent of gamma H2AX-positive GXP NS or GBM2 cells (y axis) with DMSO or TBB treatment at indicated time points of DOX treatment (x axis) is summarized in the graph in (d). Error bars represent mean±s.d. Representative 100 cells were randomly selected for quantification. Cells with ≥5 foci were considered as gamma H2AX positive. (e) Co-IF of FANCD2 & gamma H2AX with DAPI nuclear staining in TBB (25 μM/24 h)-treated GXP GBM2 exposed to DOX (0.5 μM) 5 h before fixation. Scale bars, 10 μm. The percent of FANCD2/ gamma H2AX-positive foci at DSB sites (y axis) in TBB− or TBB+ (x axis) in GXP GBM2 is summarized in the graph shown in (f). Error bars represent mean±s.e.m. See also Supplementary Fig. 4. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28094268), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - HHS-associated WRAP53 beta mutations attenuate repair of DNA double-strand breaks & lead to accumulation of DNA damage.a HeLa cells were transfected with the GFP plasmids indicated & Flag-RNF8 for 24 h; irradiated with 2 Gy; left to recover for 1 h; & then subjected to immunoprecipitation with a GFP antibody, followed by immunoblotting. b Densitometric quantifications of western blots as shown in a. The bars represent the levels of co-precipitated protein normalized to levels of the corresponding immunoprecipitated GFP protein itself, & then relative to the value obtained in wild-type WRAP53 beta . The values are means ± s.d. (the error bars) (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ns (not significant) as determined by Student’s t-test. c, d HeLa cells were transfected with siControl or siWRAP53#2 oligonucleotides for 24 h; followed by transfection with GFP-Empty or GFP-WRAP53 beta siRNA resistant plasmids for another 24 h; irradiated with 2 Gy; & 1 h later harvested for either western blotting using the antibodies indicated (c) or immunostaining for 53BP1 (d). The graph in d shows the percentage of 100 GFP-transfected cells in each experiment whose nuclei contained ≥10 53BP1 foci with the error bars depicting the s.d. (n = 4). e HeLa cells were transfected with siRNA for 8 h; followed by transfection with the GFP plasmids indicated for another 16 h; irradiated with 2 Gy; & after 24 h of recovery, immunostained for gamma H2AX. The graph shows the percentage of 100 GFP-transfected cells in each experiment whose nuclei contained ≥10 gamma H2AX foci with the error bars depicting the s.d. (n = 3). *p < 0.05, **p < 0.01, ns (not significant) as determined by Student’s t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303682), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Reptin depletion reduces the recruitment on chromatin of BRCA1 & 53BP1 after gamma ray irradiation.HuH7 cells stably expressing a doxycycline-inducible Reptin shRNA were treated with doxycycline (sh Reptin +) or left untreated (sh Reptin-) for 4 days. (A) Representative images of 53BP1 & BRCA1 foci in HuH7 cells detected using immunofluorescence 2h after gamma ray irradiation. (B) The bars represent the mean number of foci per cell from two independent experiments (>200 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, **P<0.01 & ***P<0.001, as determined by Student's t-test. (c) A2780 cells were treated as in (a) & then subjected to western blotting for WRAP53 beta , gamma H2AX, MDC1, RNF168, 53BP1, RAD51 & beta -actin. We could not assess the protein levels of BRCA1 due to a lack of antibodies that work for western blotting Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26426684), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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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NB100-926

53BP1 Antibody Pack

(11 Publications)

1 Pack

NB100-926

53BP1-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NB100-926-img0017.jpg

Immunohistochemistry: 53BP1 Antibody Pack [NB100-926] - Immunohistochemical analysis of human breast tumors stained with 53BP1 antibody (Catalog #NB100-304). Image from verified customer review.

Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - 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 Pack [NB100-926] - Detection of Human [deleted mouse] 53BP1 by Western Blot on Whole cell lysate from U2OS or 293T cells resolved on a 3 to 8% tris-acetate gel using 53BP1 Antibody (Catalog #NB100-304).

Immunohistochemistry: 53BP1 Antibody Pack [NB100-926] - Immunohistochemical staining of placental villi with 53BP1 Antibody (Catalog #NB100-305) at 40X magnification.

Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - Detection of 53BP1. 1 million Jurkat cells were fixed, permeabilized, and stained with 1.5 mcg/ml 53BP1 Antibody (Catalog #NB100-304) in a 150 mcl reaction. Isotype control (black), anti-53bp1 (red).

Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - 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.

Reactivity:

Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Nm, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Nm, Pm

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit

Host: Rabbit

Validated by:

Validated by:

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NBP2-54753

53BP1 Antibody (1285C) - BSA Free

(2 Publications)

0.1 mg

NBP2-54753

VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei." class="big_lightbox">

VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei." alt="Immunohistochemistry-Paraffin: 53BP1 Antibody (1285C) [NBP2-54753] - 53BP1 was detected in paraffin-embedded sections of human cervical cancer tissue using Rabbit Anti-Human 53BP1 Monoclonal Antibody (clone 1285C) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei." class="big_thumb" />

Western Blot: 53BP1 Antibody (1285C) [NBP2-54753] - Total protein from human A431, NTERA-2, HeLa and mouse 3T3 cell lines 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 53BP1 Antibody in block solution and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. The observed molecular weight is shown on this gel at ~250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - Neuro2a cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #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.

Flow (Intracellular): 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on HeLa Cells with 53BP1 (1285C) antibody (Catalog #NBP2-54753) (blue) and a matched isotype control MAB1050 (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - 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 [1285C] NBP2-54753 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 (1285C) [NBP2-54753] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody [1285C] NBP2-54753 (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 (1285C) [NBP2-54753] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #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.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - NIH-3T3 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #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.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - 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 [1285C] NBP2-54753 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.

53BP1 (1285C) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-53BP1 (1285C) Protein G Purified Recombinant Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NBP2-54753AF488) (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, Po, Bv, Ca, Eq, Fe, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Eq, Fe, Pm

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-54753AF350
Alexa Fluor 405	NBP2-54753AF405
Alexa Fluor 488	NBP2-54753AF488
Alexa Fluor 532	NBP2-54753AF532
Alexa Fluor 594	NBP2-54753AF594
Alexa Fluor 647	NBP2-54753AF647
Alexa Fluor 700	NBP2-54753AF700
Alexa Fluor 750	NBP2-54753AF750
Alexa Fluor Plus 405	NBP2-54753AFP405
Alexa Fluor Plus 488	NBP2-54753AFP488
Alexa Fluor Plus 555	NBP2-54753AFP555
Alexa Fluor Plus 594	NBP2-54753AFP594
Alexa Fluor Plus 647	NBP2-54753AFP647
Alexa Fluor Plus 680	NBP2-54753AFP680
Allophycocyanin	NBP2-54753APC
Biotin	NBP2-54753B
CoraFluor 1	NBP2-54753CL1
DyLight 350	NBP2-54753UV
DyLight 405	NBP2-54753V
DyLight 488	NBP2-54753G
DyLight 550	NBP2-54753R
DyLight 594	NBP2-54753DL594
DyLight 650	NBP2-54753C
DyLight 680	NBP2-54753FR
DyLight 755	NBP2-54753IR
FITC	NBP2-54753F
HRP	NBP2-54753H
Janelia Fluor 525	NBP2-54753JF525
Janelia Fluor 549	NBP2-54753JF549
Janelia Fluor 646	NBP2-54753JF646
Janelia Fluor 669	NBP2-54753JF669
PE	NBP2-54753PE
PerCP	NBP2-54753PCP
mFluor Violet 450 SE	NBP2-54753MFV450
mFluor Violet 500 SE	NBP2-54753MFV500
mFluor Violet 610 SE	NBP2-54753MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-54753SS
Azide and BSA Free	NBP2-80550

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NB100-1803

53BP1 [p Ser25] Antibody

(13 Publications)

0.1 ml

NB100-1803

Immunohistochemistry-Paraffin: 53BP1 [p Ser25] Antibody [NB100-1803] - FFPE sections of human stomach carcinoma (left) and mouse teratoma (right). Antibody: Affinity purified rabbit anti- Phospho 53BP1 (S25) used at a dilution of 1:1,000 (1ug/ml). Detection: DAB.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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AF1877

53BP1 Antibody [Unconjugated]

(1 Review)

(4 Publications)

100 μg

AF1877

HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

Western blot shows lysates of HEK293T human embryonic kidney cell line and ZR-75 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # <a class=

HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HEK293T human embryonic kidney cell line and ZR-75 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei of epithelial cells in convoluted tubules. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections." class="big_lightbox">

53BP1 was detected in perfusion fixed frozen sections of rat kidney using Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # <a class=

NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei of epithelial cells in convoluted tubules. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections." class="big_thumb" />

HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Genetic Strategies Validation. </b>Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and 53BP1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human 53BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1877) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # <a class=

HAF017). A specific band was detected for 53BP1 at approximately 350 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , ,

Applications: , , ,

Host:

Goat Polyclonal

Host: Goat 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	AF1877U-100UG
Alexa Fluor 405	AF1877V-100UG
Alexa Fluor 488	AF1877G-100UG
Alexa Fluor 532	AF1877X-100UG
Alexa Fluor 594	AF1877T-100UG
Alexa Fluor 647	AF1877R-100UG
Alexa Fluor 700	AF1877N-100UG
Alexa Fluor 750	AF1877S-100UG
Alexa Fluor Plus 405	AF1877AFP405-100UG
Alexa Fluor Plus 488	AF1877AFP488-100UG
Alexa Fluor Plus 555	AF1877AFP555-100UG
Alexa Fluor Plus 594	AF1877AFP594-100UG
Alexa Fluor Plus 647	AF1877AFP647-100UG
Alexa Fluor Plus 680	AF1877AFP680-100UG
Allophycocyanin	AF1877A
Biotin	AF1877B
CoraFluor 1	AF1877CL1
DyLight 350	AF1877D
DyLight 405	AF1877E
DyLight 488	AF1877K
DyLight 550	AF1877L
DyLight 594	AF1877M
DyLight 650	AF1877W
DyLight 680	AF1877Y
DyLight 755	AF1877Z
FITC	AF1877F
HRP	AF1877H
Janelia Fluor 525	AF1877JF525
Janelia Fluor 549	AF1877I
Janelia Fluor 646	AF1877J
Janelia Fluor 669	AF1877JF669
PE	AF1877P
PerCP	AF1877C
mFluor Violet 450 SE	AF1877MFV450
mFluor Violet 500 SE	AF1877MFV500
mFluor Violet 610 SE	AF1877MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF1877-SP

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NBP3-13664

53BP1 Antibody (HL275) - Azide and BSA Free

100 ul

NBP3-13664

Immunohistochemistry-Paraffin: 53BP1 Antibody (HL275) [NBP3-13664] - 53BP1 antibody [HL275] detects 53BP1 protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. 53BP1 stained by 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:200. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - HeLa whole cell and nuclear extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

Immunohistochemistry-Paraffin: 53BP1 Antibody (HL275) [NBP3-13664] - 53BP1 antibody [HL275] detects 53BP1 protein by immunohistochemical analysis.</br>Sample: Paraffin-embedded rat tissues.</br>53BP1 stained by 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:100.</br>Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

<b>Orthogonal Strategies Validation. </b>Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL275] diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. Corresponding RNA expression data for the same cell lines are based on Human Protein Atlas program.

Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. Corresponding RNA expression data for the same cell lines are based on Human Protein Atlas program.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, ,

Applications: , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NBP2-54677

53BP1 Antibody - BSA Free

100 ul

NBP2-54677

<b>Genetic Strategies Validation. </b>Western Blot: 53BP1 Antibody [NBP2-54677] - Western blot analysis in U-138MG cells transfected with control siRNA, target specific siRNA probe #1 and #2. Remaining relative intensity is presented. Loading control: Anti-GAPDH. The observed molecular weight as seen on this gel as greater than 250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.

ChIP-Exo-Seq composite graph for Anti-53BP1 tested in K562 cells. Strand-specific reads (blue: forward, red: reverse) and IgG controls (black: forward, grey: reverse) are plotted against the distance from a composite set of reference binding sites. The antibody exhibits robust target enrichment compared to a non-specific IgG control and precisely reveals its structural organization around the binding site. Data generated by Prof. B. F. Pugh's Lab at Cornell University.

Staining of human cerebellum shows strong nuclear positivity.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

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	NBP2-54677-25ul

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NBP3-14667

53BP1 Antibody (BL-250-1H11)

100 ul

NBP3-14667

Immunohistochemistry: 53BP1 Antibody [NBP3-14667] - Sample: FFPE section of colon carcinoma. Antibody: Rabbitanti-53BP1 recombinant monoclonal antibody used at 1:250. Secondary:HRP-conjugated goat anti-rabbit IgG.Substrate: DAB.

Immunoprecipitation: 53BP1 Antibody [NBP3-14667] - Samples: Whole cell lysate (1.0 mgper IP reaction; 20% of IP loaded) from HEK293T cellsprepared using NETN lysis buffer. Antibodies: Rabbitanti-53BP1 recombinant monoclonal antibodyused for IP at 20 ul perreaction. 53BP1 was also immunoprecipitated by rabbitanti-53BP1 antibody for blottingimmunoprecipitated 53BP1, was used at 1:1000.Chemiluminescence with an exposure time of 10 second.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Conjugate:

Unconjugated

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AF3405

53BP1 [p Ser25] Antibody

(1 Publication)

100 μg

AF3405

HAF008). A specific band for Phospho-53BP1 (S25) was detected at approximately 450 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda-phosphatase (lambda-PPase) for 1 hour. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of K562 human chronic myelogenous leukemia cell line untreated (-) or exposed (+) to 50 J/m<SUP>2</SUP>UV-C and allowed 1 hour recovery time. PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human Phospho-53BP1 (S25) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3405) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # <a class=

HAF008). A specific band for Phospho-53BP1 (S25) was detected at approximately 450 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda-phosphatase (lambda-PPase) for 1 hour. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of K562 human chronic myelogenous leukemia cell line untreated (-) or exposed (+) to 50 J/m²UV-C and allowed 1 hour recovery time. PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human Phospho-53BP1 (S25) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3405) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-53BP1 (S25) was detected at approximately 450 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda-phosphatase (lambda-PPase) for 1 hour. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

Reactivity:

Hu

Species Glossary

Reactivity: Hu

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	AF3405U-100UG
Alexa Fluor 405	AF3405V-100UG
Alexa Fluor 488	AF3405G-100UG
Alexa Fluor 532	AF3405X-100UG
Alexa Fluor 594	AF3405T-100UG
Alexa Fluor 647	AF3405R-100UG
Alexa Fluor 700	AF3405N-100UG
Alexa Fluor 750	AF3405S-100UG
Alexa Fluor Plus 405	AF3405AFP405-100UG
Alexa Fluor Plus 488	AF3405AFP488-100UG
Alexa Fluor Plus 555	AF3405AFP555-100UG
Alexa Fluor Plus 594	AF3405AFP594-100UG
Alexa Fluor Plus 647	AF3405AFP647-100UG
Alexa Fluor Plus 680	AF3405AFP680-100UG
Allophycocyanin	AF3405A
Biotin	AF3405B
CoraFluor 1	AF3405CL1
DyLight 350	AF3405D
DyLight 405	AF3405E
DyLight 488	AF3405K
DyLight 550	AF3405L
DyLight 594	AF3405M
DyLight 650	AF3405W
DyLight 680	AF3405Y
DyLight 755	AF3405Z
FITC	AF3405F
HRP	AF3405H
Janelia Fluor 525	AF3405JF525
Janelia Fluor 549	AF3405I
Janelia Fluor 646	AF3405J
Janelia Fluor 669	AF3405JF669
PE	AF3405P
PerCP	AF3405C
mFluor Violet 450 SE	AF3405MFV450
mFluor Violet 500 SE	AF3405MFV500
mFluor Violet 610 SE	AF3405MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF3405-SP

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MAB18771

53BP1 Antibody (1285A) - Azide and BSA Free

100 μg

MAB18771

Reactivity:

Hu, Po, Bv, Ca, Pm, Fe, Pm-Or, Pm, Rb, Sh

Species Glossary

Reactivity: Hu, Po, Bv, Ca, Pm, Fe, Pm-Or, Pm, Rb, Sh

Applications:

,

Applications: ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	MAB18771AF350
Alexa Fluor 405	MAB18771AF405
Alexa Fluor 488	MAB18771AF488
Alexa Fluor 532	MAB18771AF532
Alexa Fluor 594	MAB18771AF594
Alexa Fluor 647	MAB18771AF647
Alexa Fluor 700	MAB18771AF700
Alexa Fluor 750	MAB18771AF750
Alexa Fluor Plus 405	MAB18771AFP405
Alexa Fluor Plus 488	MAB18771AFP488
Alexa Fluor Plus 555	MAB18771AFP555
Alexa Fluor Plus 594	MAB18771AFP594
Alexa Fluor Plus 647	MAB18771AFP647
Alexa Fluor Plus 680	MAB18771AFP680
Biotin	MAB18771B
DyLight 350	MAB18771UV
DyLight 405	MAB18771V
DyLight 488	MAB18771G
DyLight 550	MAB18771R
DyLight 594	MAB18771DL594
DyLight 650	MAB18771C
DyLight 680	MAB18771FR
DyLight 755	MAB18771IR
FITC	MAB18771F
HRP	MAB18771H
Janelia Fluor 549	MAB18771JF549
Janelia Fluor 646	MAB18771JF646

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	MAB18771-SP

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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

Modification

Clonality

Clone

Images Type

Isotype

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

TP53BP1 Products By Gene Name

53BP1 Antibody - BSA Free

53BP1 Antibody - BSA Free

53BP1 Antibody - BSA Free

53BP1 Antibody Pack

53BP1 Antibody (1285C) - BSA Free

53BP1 [p Ser25] Antibody

53BP1 Antibody [Unconjugated]

53BP1 Antibody (HL275) - Azide and BSA Free

53BP1 Antibody - BSA Free

53BP1 Antibody (BL-250-1H11)

53BP1 [p Ser25] Antibody

53BP1 Antibody (1285A) - Azide and BSA Free