Product Search for: Novus Biologicals

NB100-384

Histone H2AX [p Ser139] Antibody - BSA Free

(1 Review)

(180 Publications)

0.1 ml

NB100-384

Detection of Human and Mouse Histone H2AX [p Ser139] by Western Blot. Samples: Nuclear extract (50 ug) from human HEK293, human melanoma (G361), mouse wildtype embryonic fibroblasts (+/+) or mouse H2AX knockout embryonic fibroblasts (-/-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody NB100-384 used at 0.1 ug/ml. Detection: Chemiluminescence with 30 second exposure. (NCS, neocarzinostatin - 200 ng/ml, 30 min). Bands appear at an observed molecular weight of ~15 kDa.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Samples: Neocarzinostatin treated asynchronous HeLa cells (left) and untreated asynchronous HeLa cells (right) . Antibody: Affinity purified rabbit Histone H2AX [p Ser139] used at a dilution of 1:5,000 (0.2ug/ml). Detection: Red fluorescent Anti-rabbit IgG-DyLight 594 used at a dilution of 1:100.

Simple Western lane view shows a specific band for Histone H2AX [p Ser139] in 0.2 mg/ml of Jurkat lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa separation system.

<b>Biological Strategies Validation. </b>Immunocompromised mice were subcutaneously injected with cancerous cell lines and tumors were allowed to establish. Treatments occurred every other day and the studied compound or the equivalent vehicle control administered intraperitoneally for five weeks. Tumor volume and mass were measured two times per week. IHC analysis of sectioned tumor tissues from the MDA-MB-231 study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41598-017-01230-4), licensed under a CC-BY license.

Immunostaining of Histone H2AX [p Ser139] WT1 and 5mC in patients with IgA nephropathy and controls. Examples of PAS staining and immunostaining with Histone H2AX [p Ser139] (green) and WT1 (red), pATM and 5mC in glomeruli of IgA nephropathy and controls. A kidney sample of a 65-year-old male of IgA nephropathy without podocytopathic features. Arrows indicate Histone H2AX [p Ser139] and WT1 double-positive cells. Scale bars: 50um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-57140-0) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Samples: Nuclear extract from HeLa cells treated with 100 uM EPE for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes. Band appears at an observed molecular weight of ~17 kDa.

<b>Biological Strategies Validation. </b>Detection of human Histone H2AX [p Ser139] by western blot. Samples: Whole cell lysate (50 ug) from Jurkat cells treated with 100 uM EPE for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody NB100-384 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 seconds. Band appears at an observed molecular weight of ~18 kDa.

FFPE section of mouse CT26 colon carcinoma. Antibody: Affinity purified rabbi Histone H2AX [p Ser139] antibody used at a dilution of 1:1,000 (1 ug/ml). Detection: DAB.

Analysis of Histone H2AX [p Ser139] in EPE Treated Jurkat Cells. Cells were treated for 3 hrs in 5ug/ml etoposide, fixed in 1.5% PFA, and permeabilized in 90% Methanol. 1 million cells were stained with 0.5 ug anti-KLH or anti-H2AX NB100-384 and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- etosposide treated, anti-KLH; Red- untreated, anti-Histone H2AX [p Ser139]; Blue- etoposide treated , anti-Histone H2AX [p Ser139].

FFPE section of human ovarian carcinoma. Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at a dilution of 1:5,000 (0.2 ug/ml). Detection: DAB.

Electropherogram image(s) of corresponding Simple Western lane view. Histone H2AX [p Ser139] antibody was used at 5 ug/ml dilution on Jurkat lysate(s).

Immunohistochemistry: Rabbit Polyclonal Histone H2AX [p Ser139] Antibody - Histone H2AX Antibody on mouse cancer tissue. H2AX (Gray) and H2BGFP(Green). Primary antibody dilution: 1:1000 in a 10um slice.

mTORC1/2 activity prevents Cisplatin-induced cell death in MCF-10A cells. (A) Western blot displaying effects on mTOR signaling during a dose escalation of PP242 treatment in MCF-10A cells; (B) Western blot displaying effects of mTOR signaling on a dose escalation of cisplatin treatment in MCF-10A cells; (C) Western blot displaying effects on mTOR signaling and cell death during non-treated, Cisplatin, PP242, and Cisplatin + PP242-treated MCF-10A cells.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - ATM & NF-kappa B activation are downregulated in Ercc1-/ delta mice heterozygous for Atm. (A) Livers were collected at 12 weeks of age from WT, Ercc1-/ delta & Ercc1-/ delta Atm+/- mic (n=3 per genotype) & lysates analyzed by western blot for activation of ATM & its downstream effectors. (B) Same liver lysates were used to measure phosphorylation of p65 & I kappa B alpha . (C) Western blot analysis of livers from 16-week-old WT, Ercc1-/ delta & Ercc1-/ delta Atm+/-mice (n=3 per genotype) probed for activation of ATM. GAPDH was used as a loading control. (D) Same liver lysates used to measure activation of NF-kappa B. (E) Fourteen-week-old livers from Ercc1-/ delta & Ercc1-/ delta p65+/- mice (n=3 per genotype) were analyzed by western blot for activation of ATM (F) & NF-kappa B. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - Immunostaining of gamma H2AX, WT1 & 5mC in patients with IgA nephropathy & controls. Examples of PAS staining & immunostaining with gamma H2AX (green) & WT1 (red), pATM & 5mC in glomeruli of IgA nephropathy & controls. (A) A control kidney sample of 44-year-old female, (B) 65-year-old male of IgA nephropathy without podocytopathic features & (C) 55-year-old male of IgA nephropathy with podocytopathic features. Arrows indicate gamma H2AX & WT1 double-positive cells. Scale bars: 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31937846), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - UHRF1 deficiency resulted in impaired meiotic recombination & defective pachynema.a Double immunofluorescence of SYCP3 (green) & DMC1 (red) in testicular spread preparations. b, c The number of DMC1 foci in zygotene stage (b) & pachytene stage (c). d Immunostaining for SYCP3 (red) & gamma H2AX (green). e The percentage of abnormal gamma H2AX foci in the pachytene stage. f Immunostaining for SYCP3 (red) & MLH1 (green). g The number of MLH1 foci in pachynema. h Immunostaining for SYCP3 (red) & H1t (green). i The percentage of spermatocytes with H1T staining. ***p ≤ 0.001; *p ≤ 0.05. Scale bar, 5 μm in a, d, f, h. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32081844), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - mTORC1/2 activity prevents Cisplatin-induced cell death in MCF-10A cells. (A) Western blot displaying effects on mTOR signaling during a dose escalation of PP242 treatment in MCF-10A cells; (B) Western blot displaying effects of mTOR signaling on a dose escalation of cisplatin treatment in MCF-10A cells; (C) Western blot displaying effects on mTOR signaling & cell death during non-treated, Cisplatin, PP242, & Cisplatin + PP242-treated MCF-10A cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31771139), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Pharmacologic inhibition of ATM rescues oxidative stress-induced senescence by suppressing ATM- & NEMO-mediated NF-kappa B activation. (A) Representative images of primary WT & Ercc1-/- MEFs were induced to undergo senescence by serial passaging at 20% oxygen. At passage 5, MEFs were grown in the presence or absence of KU-55933 (10 μM) for 72 hrs. Senescence was determined by SA-beta gal staining. Images were obtained at the magnification of 10x. (B) Quantitation of the percent SA-beta gal positive cells. Graph represents the mean +/- s.e.m. of three independent experiments. Student’s t-test, ***p <0.001, ****p <0.0001. (C) Passage 5 Ercc1-/- MEFs treated with vehicle or KU-55933 (10 μM) for 72 hours were collected & levels of p21 & p16INK4a were determined by western blotting. (D) Passage 5 Ercc1-/- MEFs were treated with KU-55933 (10 μM) for 72 hours & whole cell lysate (CL) & nuclear extracts (NE) were analyzed by immunoblotting for expression of proteins involved in the DNA damage response. (E) Whole cell lysate (CL) & nuclear extract (NE) were extracted from Ercc1-/- MEFs treated with 10 μM of KU-55933 for analysis of nuclear NEMO & p65. GAPDH was used as a loading control of total proteins & LaminA/C as a loading control of nuclear protein. (F) Passage 5 WT & Ercc1-/- MEFs transfected with a NF-kappa B-luciferase reporter construct were cultured in the presence or absence of KU-55933 (10 μM) & were collected for luciferase assays after 72 hours. (G) qRT-PCR analysis of mRNA expression in passage 5 WT & Ercc1-/- MEFs treated with or without of KU-55933 (10 μM) for 72 hrs. P values were determined using a Student’s t-test. *p<0.05, **p<0.01, ***p <0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-384] - Orally administered lemongrass & white tea extract reduce tumor size in lymphoma xenograft model in immunocompromised miceImmunocompromised mice were subcutaneously injected with cancerous cells & tumors were allowed to establish. Treatments occurred every other day & the studied compound or the equivalent vehicle control administered orally for three weeks. (A, B) The tumors were photographed before & after extraction from the animals. (C) Tumor volume & mass were measured two times per week. (D) Immunohistochemistry analysis of sectioned tumor tissues from the lymphoma study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Statistical analysis using One-Way ANOVA. *p < 0.05 vs tumor volume of the control. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.22502), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - CPT induces transcription- & proteasome-dependent apoptosis in quiescent WI38 hTERT cells. (A & B) Serum-starved cells were treated with DMSO or FLV (1 μM) for 1 h before the addition of DMSO or CPT (25 μM) for 16 & 24 h. (A) Percentages of cells that remained attached to culture flasks (means ± SD of quadruplicates). (B) Cell survival was analyzed by a CellTiter-Blue assay (means ± SD of triplicates). (C–G) Western blot of the indicated proteins in serum-starved cells treated for 1 h with DMSO or with FLV (1 μM) (C), lactacystin (10 μM) (D), veliparib (5 μM) or olaparib (10 μM) (E), ATMi (10 μM) (F) or DNA-PKi (10 μM) (G) before the addition of DMSO (‘-’, 24 h in panels C & F) or CPT (25 μM) for the indicated times. Data shown are representatives from two to three experiments. PARPCL: cleaved PARP. H2AX & alpha Tubulin were the loading controls. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Inhibition of DNA-PK prevents monoubiquitination of H2AX & H2A in CPT-treated quiescent WI38 hTERT cells. (A–E) Serum-starved cells were treated with DMSO or DNA-PKi (10 μM) for 1 h before the addition of DMSO (untreated) or CPT (25 μM) for 1 h. (A) Western blot of gamma H2AX. +Ub1 indicates gamma H2AX monoubiquitinated. The top panel shows quantification of Ub1-gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4). **P < 0.01. (B & C) Cells were pre-extracted with CSK buffer before co-staining for Ub-H2A (red) & 53BP1 phosphorylated on S1778 (p53BP1) (green). (B) Representative pictures. Images were merged to determine colocalization (yellow). The large Ub-H2AX focus at the periphery of the nuclei of untreated & CPT-treated cells may marks the inactive X chromosome as reported (91). (C) Percentages of nuclei with at least 5 Ub-H2A foci (means ± SEM, n = 3, 100 nuclei were analyzed for each treatment in each experiment). ***P < 0.001. (D & E) Cells were co-stained for ubiquitinated proteins (FK2, red) & gamma H2AX (green). (D) Representative pictures. Images were merged to determine colocalization (yellow). (E) Number of FK2 foci per nucleus from one representative experiment (76–111 nuclei were analyzed for each treatment) out of three. ****P < 0.0001. In the microscopic images, nuclear contours, identified by DAPI staining (blue in the merge images at bottom), are indicated by dashed lines. Bars: 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - 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 & cullin 4B or against a control sequence & then treated with DMSO (−CPT) or 25 μM CPT (+CPT) for 1 h. (A & 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 & 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) & 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 & 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) & 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) & 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 & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - Inhibition of DNA-PK prevents monoubiquitination of H2AX & H2A in CPT-treated quiescent WI38 hTERT cells. (A–E) Serum-starved cells were treated with DMSO or DNA-PKi (10 μM) for 1 h before the addition of DMSO (untreated) or CPT (25 μM) for 1 h. (A) Western blot of gamma H2AX. +Ub1 indicates gamma H2AX monoubiquitinated. The top panel shows quantification of Ub1-gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4). **P < 0.01. (B & C) Cells were pre-extracted with CSK buffer before co-staining for Ub-H2A (red) & 53BP1 phosphorylated on S1778 (p53BP1) (green). (B) Representative pictures. Images were merged to determine colocalization (yellow). The large Ub-H2AX focus at the periphery of the nuclei of untreated & CPT-treated cells may marks the inactive X chromosome as reported (91). (C) Percentages of nuclei with at least 5 Ub-H2A foci (means ± SEM, n = 3, 100 nuclei were analyzed for each treatment in each experiment). ***P < 0.001. (D & E) Cells were co-stained for ubiquitinated proteins (FK2, red) & gamma H2AX (green). (D) Representative pictures. Images were merged to determine colocalization (yellow). (E) Number of FK2 foci per nucleus from one representative experiment (76–111 nuclei were analyzed for each treatment) out of three. ****P < 0.0001. In the microscopic images, nuclear contours, identified by DAPI staining (blue in the merge images at bottom), are indicated by dashed lines. Bars: 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - DDR & NF-kappa B are activated concomitantly in senescent MEFs & aged tissues. (A) Immunoblot detection of p-p65 & total p65 in liver tissue from 16-week-old WT (n=3) & Ercc1-/ delta (n=3) mice. (B) Immunoblot detection of phosphorylation of ATM & downstream targets gamma H2AX & p21 in liver from 16-week-old WT & Ercc1-/ delta mice. (C) Immunoblot detection of phosphorylation of NF-kappa B & I kappa B alpha in liver lysates from 3, 12 & 24 month-old WT mice. n=3 mice per group. (D) Immunoblot detection of p-ATM, ATM & p21 in the same liver lysates. (E) Immunoblot detection of DDR effectors in nuclear extracts from passage 5 WT & Ercc1-/- MEFs, grown at 20% oxygen. (F) Level of NF-kappa B activation is higher in Ercc1-/- MEFs compared to WT MEFs at passage 5, as measured by Immunoblot detection of p-p65 & total p65 in WT & Ercc1-/- MEFs at passage 5 after culturing in 20% oxygen. (G) Representative images of immunofluorescent detection of p65 & NEMO in passage 4 WT & Ercc1-/- MEFs grown at 20% oxygen. Blue: DAPI staining; Green: p65 (top panel) or NEMO (bottom panel). Images were taken at the magnification of 60x. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Induction of ubiquitin/proteasome-dependent DSBs in CPT-treated quiescent WI38 hTERT cells. (A–F) Serum-starved cells were treated with DMSO (1 h) or with MG132 (50 μM, 1 h), lactacystin (10 μM, 1 h), bortezomib (1 μM, 4 h), G5 (1.5 μM, 0.5 h) or Pyr-41 (9 μM, 0.5 h) before the addition of DMSO (untreated) or 25 μM CPT for 1 h & then co-stained for gamma H2AX (green) & 53BP1 (red) or analyzed by Western blot. ‘-’ in panels C & F means cells treated with DMSO. (A & D) Representative pictures. Nuclear contours, identified by DAPI staining (not shown), are indicated by dashed lines. Bars: 10 μm. (B & E) Number of gamma H2AX foci per nucleus from two independent experiments (147–153 nuclei were analyzed for each treatment). ****P < 0.0001. (C & F) Western blot of gamma H2AX. alpha Tubulin: loading control. Dashed lines indicate that intervening wells have been spliced out. The top panels show quantification of gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4 in panel C, n = 3 in panel F). ***P < 0.001; **P < 0.01. (G & H) Detection of DSBs by neutral Comet assays in serum-starved cells treated with DMSO or MG132 (25 μM) for 1 h before the addition of DMSO (untreated) or CPT for 1 h (7.5 μM for experiments (Exp) I & II; 5 & 7.5 μM for Exp III). (G) Representative pictures of nuclei from Exp I. (H) Quantification of neutral Comet tail moments for three independent experiments (95–133 nuclei were analyzed for each treatment in each experiment). ***P < 0.001; ****P < 0.0001. The untreated & CPT data from Exp I are from the same experiment as that of Supplementary Figure S3D. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Curcumin Analogs Induce Apoptosis in Cancerous Cells by Several Pathways. (a) E6-1 (Jurkat), dominant negative FADD (dnFADD) Jurkat, & overexpressing BCL-2 Jurkat were treated for 48 hours then stained for Annexin V & PI (b) E6-1 cells were plated & treated with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK for 48 hours. Cells were stained for Annexin V & PI. Results were obtained using image-based cytometry with the Y-axis representative of percent of cells positive for Annexin V (green), PI (red), Annexin V & PI (yellow), or negative for both Annexin V & PI (blue). Values are expressed as a mean ± SD from three independent experiments. (c) E6-1 cells were treated for 24 hours with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK & the studied compounds, lysed & subjected to Western blot analysis. (d) NHF & NCM460 cells were treated for 48 hours & 72 hours respectively, lysed & subjected to Western blot analysis. Bands were visualized with a chemiluminescence reagent. Images are representative of three independent experiments. Statistical calculations were performed using Two-Way ANOVA multiple comparison. *p < 0.05 vs % viable of Control (DMSO); #p < 0.05 vs % viable cells for groups without Z-VAD(oMe)FMK. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28439094), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Ca

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-384AF350
Alexa Fluor 405	NB100-384AF405
Alexa Fluor 488	NB100-384AF488
Alexa Fluor 532	NB100-384AF532
Alexa Fluor 594	NB100-384AF594
Alexa Fluor 647	NB100-384AF647
Alexa Fluor 700	NB100-384AF700
Alexa Fluor 750	NB100-384AF750
Allophycocyanin	NB100-384APC
Biotin	NB100-384B
CoraFluor 1	NB100-384CL1
DyLight 350	NB100-384UV
DyLight 405	NB100-384V
DyLight 488	NB100-384G
DyLight 550	NB100-384R
DyLight 594	NB100-384DL594
DyLight 650	NB100-384C
DyLight 680	NB100-384FR
DyLight 755	NB100-384IR
FITC	NB100-384F
HRP	NB100-384H
Janelia Fluor 525	NB100-384JF525
Janelia Fluor 549	NB100-384JF549
Janelia Fluor 646	NB100-384JF646
Janelia Fluor 669	NB100-384JF669
PE	NB100-384PE
PerCP	NB100-384PCP
mFluor Violet 450 SE	NB100-384MFV450
mFluor Violet 500 SE	NB100-384MFV500
mFluor Violet 610 SE	NB100-384MFV610

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AB-105-C

Rabbit IgG Isotype Control [Unconjugated]

(2 Reviews)

(178 Publications)

1 mg

AB-105-C

HepG2 human hepatocellular carcinoma cell line was stained with Rabbit Anti-Human Klotho beta Monoclonal Antibody (Catalog # MAB58891, filled histogram) or Rabbit IgG Isotype Control Antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). View our protocol for Staining Membrane-associated Proteins.

<b>Biological Strategies Validation. </b>pSTAT5, but not NS1, interacts with the MCM complex.(A) Immunoprecipitation (IP) assay. Cell lysates of NS1Flag-expressing UT7/Epo-S1 cells were prepared for pull-down assays with either anti-Flag-conjugated beads or control beads. Immunoprecipitated proteins were examined for the presence of MCM2 by Western blotting. Blots were reprobed with rabbit anti-pSTAT5(Y694), anti-E2F5, and anti-Flag antibodies. Detection of E2F5 was used as a positive control for NS1 IP. (B) Co-IP assay. UT7/Epo-S1 cells were collected, washed, and lysed with RIPA buffer. After centrifugation, the supernatant was incubated with either rabbit anti-pSTAT5(Y694) or control IgG antibody. Immunoprecipitated proteins were blotted for the presence of the MCM complex with an anti-MCM5 antibody and for pSTAT5 with rabbit anti-pSTAT5(Y694). (C) Reverse Co-IP assay. Reverse Co-IP was performed with an anti-MCM5 antibody. Immunoprecipitated proteins were examined for pSTAT5, MCM2, and MCM5, respectively. (D) Co-IP of lysates treated with DNase. UT7/Epo-S1 cell lysates, either treated or untreated with DNase (750 units of Benzonase) were incubated with anti-pSTAT5(Y694) or control IgG antibodies for Co-IP assay, and immunoprecipitated proteins were examined for MCM2 by Western blot analysis. (E-H) Immunofluorescence analysis. (E&F) Mock- or B19V-infected CD36+ EPCs were co-stained with rabbit anti-STAT5 and mouse anti-MCM2 antibodies, followed by (E) incubation with respective secondary antibodies, or by (F) proximal ligation assay, which produces amplified signal for labeled molecules in close proximity. (G) CD36+ EPCs were incubated with either DMSO or pimozide (at 30 μM) for 2 days. And then the cells were co-stained with rabbit anti-STAT5 and mouse anti-MCM2 antibodies for proximity ligation assay. (H) Infected EPCs were stained with an anti-capsid antibody. Confocal images were taken with an Eclipse C1 Plus (Nikon) microscope at 100 × magnification. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1006370), licensed under a CC-BY license. Not internally tested by R&D Systems.

Elevated TGF-beta levels in the early inflammatory stage of AS. a Hematoxylin and eosin (H&E) staining and b Safranin O–Fast Green (SOFG) staining of normal and inflamed interspinous ligaments. In the AS group, the right panels show magnified views of the boxed area in the left panels. Scale bar: 100 μm (two panels on the right); 25 μm (left panel). c Immunostaining and d quantitative analysis of CD68-positive cells (brown) in the normal and inflamed interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). e Immunostaining and f quantitative analysis of pSmad2/3-positive cells (brown) in the normal and inflamed interspinous ligaments (sagittal view) in normal ligaments and inflammatory ligaments. The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). g Immunostaining and h quantitative analysis of pSmad1/5/8-positive cells (brown) in the normal and inflamed interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Osteoclast resorption of bony interspinous ligaments releases active TGF-beta to drive the progression of ossification in AS patients. e Immunostaining and f quantitative analysis of CD68-positive cells (brown) in the normal and HO-formed interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Chondrocyte differentiation and cartilage formation in interspinous ligaments of AS patients before calcification. a Hematoxylin and eosin (H&E) staining and b Safranin O–Fast Green (SOFG) staining of normal and chondrogenic interspinous ligaments. In the AS group, the right panels show magnified views of the boxed area in the left panels. Scale bar: 100 μm (two panels on the right side); 25 μm (left panel). c Immunostaining and d quantitative analysis of collagen II-positive cells (brown) in the normal and chondrogenic interspinous ligaments (sagittal view). The bottom panels show a magnified view of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). e Immunostaining and f quantitative analysis of pSmad2/3-positive cells (brown) in the normal and chondrogenic interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). g Immunostaining and h quantitative analysis of pSmad1/5/8-positive cells (brown) in the normal and inflammatory interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). C, cartilage; L, ligament Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Progression of endochondral heterotopic ossification in AS patients. i Immunostaining and j quantitative analysis of pSmad2/3-positive cells (brown) in the normal ligaments and endochondral-ossified interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). k Immunostaining and l quantitative analysis of Osterix-positive cells (brown) in the normal and endochondral-ossified interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). m Immunostaining and n quantitative analysis of pSmad1/5/8-positive cells (brown) in the normal and inflammatory interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). B, bone; BM, bone marrow; C, cartilage Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Progression of endochondral heterotopic ossification in AS patients. g Immunostaining and h quantitative analysis of the number of CD68-positive osteoclast (brown) surface (OCS) per bone surface (BS). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). i Immunostaining and j quantitative analysis of pSmad2/3-positive cells (brown) in the normal ligaments and endochondral-ossified interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). k Immunostaining and l quantitative analysis of Osterix-positive cells (brown) in the normal and endochondral-ossified interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). m Immunostaining and n quantitative analysis of pSmad1/5/8-positive cells (brown) in the normal and inflammatory interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). B, bone; BM, bone marrow; C, cartilage Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Osteoclast resorption of bony interspinous ligaments releases active TGF-beta to drive the progression of ossification in AS patients. g Immunostaining and h quantitative analysis of pSmad2/3-positive cells in the normal and HO-formed interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). i Immunostaining and j quantitative analysis of Osterix-positive cells (brown) in the normal and HO-formed interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panel); 25 μm (bottom panel). k Immunostaining and l quantitative analysis of PDGF-BB-positive cells in the normal and HO-formed interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). m CD31-positive (red) cells, n EMCN-positive (green) cells and quantification of the fold change of o CD31-positive vessels, p EMCN-positive vessels in the normal and HO-formed interspinous ligaments (sagittal view). q PGP9.5-positive (red) cells and CGRP-positive (green) cells and quantification of the fold change of r PGP9.5 and CGRP double positive nerves in the normal and HO-formed interspinous ligaments (sagittal view). s Immunostaining and t quantitative analysis of pSmad1/5/8-positive cells (brown) in the normal and inflammatory interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Progression of endochondral heterotopic ossification in AS patients. c Immunostaining and d quantitative analysis of collagen II-positive cells (brown) in the normal and chondrogenic interspinous ligaments (sagittal view). The bottom panels show magnified views of the boxed area in the top panels. Scale bar: 100 μm (top panels); 25 μm (bottom panels). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33731675), licensed under a CC-BY license. Not internally tested by R&D Systems.

$Protein-protein interaction between ANO1 and NCX3. (A) Co- IP of ANO1 and NCX3 from small intestine smooth muscle membrane fraction. Immunoprecipitates (IP) were obtained by elution with low pH buffer, and immunoblotting (IB) was performed using Wes with anti-ANO1 and anti-NCX3 antibodies. Lanes: 1, protein standards; 2, ANO1 in membrane fraction (2.5 mg); 3, ANO1 in ANO1 immunoprecipitate (5 ml); 4, ANO1 in non-immune rabbit IgG IP; 5, NCX3 in membrane fraction (1 mg); 6, NCX3 in NCX3 immunoprecipitate (5 ml); 7, NCX3 in non-immune rabbit IgG IP; 8, NCX3 in Ano1 immunoprecipitate (5 ml); 9, ANO1 in NCX3 immunoprecipitate (5 ml) n = 4. (B) Association between ANO1 and NCX3 occurs in situ in ICC, as demonstrated by PLA. ICC from small intestine (green, left panel) were exposed to the rabbit anti-ANO1 and goat anti-NCX3 antibodies, followed by the Duolink minus-anti rabbit IgG and plus-anti goat IgG secondary antibodies and the ligation and amplification reactions. For the PLA negative control, ICC were exposed to rabbit anti-ANO1 and mouse anti-myosin regulatory light chain antibodies (upper left panel). PLA-positive red spots were detected by ANO1-NCX3 co-localization (lower left panel). Few red spots were observed in negative controls. Nuclear staining of ICC was achieved with DAPI (blue, left panel). Intact ICC were confirmed by differential interference contrast (DIC) (right panels). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32256387), licensed under a CC-BY license. Not internally tested by R&D Systems.$

Reactivity:

Rb

Species Glossary

Reactivity: Rb

Applications:

Applications:

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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Validated by:

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NB100-2284

CD31/PECAM-1 Antibody

(5 Reviews)

(151 Publications)

0.1 ml

NB100-2284

Staining of CD31/PECAM-1 in Rat Liver sample. IHC-P image submitted by a verified customer review.

Human lung adenocarcinoma using. Antibody at 1:100 with citrate epitope retrieval at pH 6.0.

Analysis in rat muscle blood vessels using. IHC-P image submitted by a verifed customer review.

Matrix-HA supplemented with strontium was implanted subcutaneously in mice, where immunochemistry analysis and histology occurred of the newly formed tissues. (C) Within each matrix: Matrix-HA, Matrix-8Sr-HA and Matrix-50Sr-HA, were indicated by immunostaining of CD31 for the newly formed tissues. (D) Number of vessels within each tissue was quantified using NDP view software. Unit is square mm. Immunostaining analysis of slides occurred for 2 samples per condition and 3 sections were analyzed per sample and per group of matrix. Citation: Ehret C, Aid-Launais R, Sagardoy T, Siadous R, Bareille R, Rey S, et al. (2017) Strontium-doped hydroxyapatite polysaccharide materials effect on ectopic bone formation. PLoS ONE 12(9): e0184663. //doi.org/10.1371/journal.pone.0184663

Imaging of HUVEC monolayer on glass substrate. Fixation with 4% PFA. Blocking with Goat Serum at 1:100. Incubation 2 hours at room temperature. ICC/IF image submitted by a verified customer review.

Analysis of canine kidney tissue using CD31/PECAM-1 antibody. Antibody was used at a 1:50 concentration in Casein in PBS and left at 4C overnight on paraffin embedded canine bladder tissue. HIER was performed in Tris/EDTA buffer, pH 9 for two hours at 75C. Image from verified customer review.

Analysis in pig SC cells using CD31/PECAM-1 Antibody (red). Blue color showing nucleus labeling. ICC/IF image submitted by a verified customer review.

Immunohistochemistry evaluation of CD31/PECAM-1 expression. Representative images of CD31/PECAM-1 staining of vessels in the three groups of implanted matrices (scale bar = 100 um). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-06258-0) licensed under a CC-BY license.

CD31 staining on E11.5 mouse Pharyngeal Mesoderm. Fixed with 4% PFA overnight. Blocked with 1% BSA. Primary antibody at 1:100. Secondary antibody at 1:1000, conjugated to Alexa Fluor 488. IHC-Fr image submitted by a verfied customer review.

Immunohistochemistry evaluation of CD31 expression. (a) Representative images of CD31 staining of vessels in the three groups of implanted matrices (scale bar = 100 µm). (b) Quantification of vessel density inside the implanted matrices at 5 weeks post implantation (n = 6; Average ± SD). NS and **denote Non Significant and p < 0.01, respectively.

Immunohistochemistry: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - Staining of CD31 in human lung tissue. Nuclei in blue and CD31 (vessel) in green. Mouse anti-CD31 (1:100, overnight incubation at +4°C), followed by donkey anti-mouse 488 (1:400, 3h at RT). Image from a verified customer review.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Antibodies selected for characterization of stromal cells & vasculature in FFPE specimens are species-specific. (A, B) Labeling of a patient colon tumor (A) & Colo205 human cell line xenograft (B) with anti-human mitochondrial antibody (single arrow- tumor cells; double arrows- stroma). (C, D) Labeling of a patient pancreatic tumor (C) & Colo205 human cell line xenograft (D) with anti-human CD34. (E, F) Labeling of a patient lung tumor (E) & Colo205 xenograft (F) with anti-human CD31. (G, H) Labeling of a patient lung tumor (G) & Colo205 xenograft (H) with anti-mouse CD34. (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - At the time of second passage, vessels in successfully growing xenografts of four different tumor types surveyed were of murine origin: A-C Colon; D-F Lung; G-I Pancreatic; J-L Renal Cell Carcinoma. For each tumor type, a representative section of an original patient specimen labeled with anti-huCD31 is shown (A, D, G, J). For each tumor, sections of the first passage xenografted tumor when it was resected are also shown; while no huCD31(+) vessels were identified in the xenografts (B, E, H, K), msCD34(+) vessels were abundant (C, F, I, L). (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Vascularization of an engrafted patient colon tumor. A patient colon tumor was implanted in a cohort of mice & vessel development was analyzed over 8 weeks. Vessels in the original patient specimen labeled strongly for huCD31 (A) & not for msCD34 (D). Representative sections showing loss of huCD31(+) vessels (B- 4 weeks, C- 7 weeks) & presence of msCD34(+) vessels (E- 4 weeks, F- 7 weeks) are shown. The graph (G) summarizes this process; huCD31(+) vessels were rapidly lost, & by one week, mouse vessels were the predominate vessels present in the colon tumors (no data for msCD34 at 2 weeks; bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: CD31/PECAM-1 Antibody [NB100-2284] - KCl pulse decreased membrane, but increased nuclear, detection of NHE1 in b.End3 endothelial cells.(A) Representative immunofluorescence images of bEnd.3 cells at two time-points after KCl or aCSF pulse. (B) Representative immunoblots of NHE1, alpha -tubulin, PECAM, & lamin B in cytosol, membrane, & nuclear fractions of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. (C = cytosol, M = membrane, N = nuclear) Values represent the mean ratio of NHE1 detection ± SEM (n = 11–12). (C) Representative immunoblots indicating NHE1 & alpha -tubulin as a loading control in whole cell lysate of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. Values represent the % of aCSF-treated relative expression ± SEM (n = 6). (D) Representative immunoblots of NHE1 & alpha -tubulin as a loading control in whole lysate of microvessels harvested at 90 min after cortical injection of KCl or aCSF. Values represent the % of naive relative expression ± SEM (n = 6). # denotes significantly different vs naïve (p<0.01), as assessed by one-way ANOVA (E) Intracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 45). (F) Extracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 6) in triplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32469979), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - Impact of T‐AuNPs on endothelial cells in vivo. Immunofluorescent staining for endothelial cells using an anti‐CD 31 antibody revealed an intact endothelial cell monolayer in the uninjured control carotid artery (N = 3). As expected post balloon angioplasty, there is no endothelial cell monolayer of the injured left carotid arteries in the injury alone, injury + T‐AuNP, & injury + S‐AuNP treatment conditions. Green indicates autofluorescence of the elastic lamina. Magenta indicates endothelial cells. Images obtained using 25× magnification with exposure time of 400 msec. TUNEL staining for apoptosis revealed no evidence of endothelial cell apoptosis in the uninjured control carotid arteries, with a prominent endothelial cell monolayer (white arrowhead). Mild apoptosis was noted in the media of balloon‐injured left carotid arteries, as expected. Black arrows indicate apoptotic cells. Representative images obtained using 40× magnification. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28242820), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - The growth of a patient mesothelioma xenograft was supported by development of a murine vascular network. A patient mesothelioma was implanted in a cohort of mice & monitored for tumor growth. Once tumors began to actively grow, representative tumors were resected at weekly intervals & analyzed for vessel content. Staining for huCD31 was prominent in the original patient specimen (A), much reduced at 4 weeks (B) & negligible at 9 weeks (C). In contrast, patient specimens were not stained for msCD34 (D), whereas at 4 weeks large numbers of vessels stained for msCD34 (E) & by 9 weeks, msCD34 labeled vessels were predominant (F). The graph (G) summarizes the loss of detectable human vessels & acquisition of murine vessels over a 9 week period. (bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - TGF‐ beta 1 released by MSCs contributes to induced myofibroblast differentiation & granulation tissue formationA–C2.5 × 105 of TGF‐ beta 1 siRNA or control siRNA‐transfected AT‐MSCs were intradermally injected around each of CD18−/− murine wound. PBS mock injection served as negative control. Wound tissue was harvested at day 2, 5, & 7 post‐wounding for quantification of human TGF‐ beta 1 mRNA (A) at day 2 by qPCR, total TGF‐ beta 1 (B), & active TGF‐ beta 1 (C) protein at day 5 by ELISA. Data are expressed as mean ± SEM, n = 3 wounds per group, **P < 0.01, by one‐way ANOVA with Tukey's test.D–GExpression of alpha ‐SMA (D & E) & CD31 (F & G) at days 5 & 7 by immunostaining on tissue sections. The dashed lines indicate the border of the wound bed & epidermis or eschar. e, epidermis; es, eschar; wb, wound bed. Scale bars: 100 μm. Quantification data are expressed as mean ± SEM, n = 3 wounds per group, *P < 0.05, ***P < 0.001, by one‐way ANOVA with Tukey's test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32080965), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - CD31 positive vessels in a canine splenic hemangiosarcoma.Tissue slides were loaded into automated research stainer, dewaxed and pretreated with EDTA-based epitope retrieval ER2 solution for 20 mins at 100°C. After a peroxide incubation for 5 min, the rabbit antibody against CD31 (NB100-2284) (1:100) was incubated for 60 mins at room temperature followed by Bond Polymer (anti-rabbit HRP) incubation for 8 min at room temperature. Mixed DAB reagent (Polymer Refine Detection Kit) was incubated for 10 mins, and Hematoxylin (Refine Detection Kit) counterstaining for 10 mins. After staining, sample slides were washed in water, dehydrated using ethanol gradient (70%, 90%, 100%), washed three times, and mounted in mounting medium. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Po, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

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Unconjugated

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NB300-141

GFAP Antibody

(12 Reviews)

(117 Publications)

0.05 ml

NB300-141

Simple Western: GFAP Antibody [NB300-141] - Simple Western lane view shows a specific band for GFAP in 0.05 mg/mL of Human Brain lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: GFAP Antibody [NB300-141] - The inhibition of Kv1.3 channels induces neuroprotection against the toxic effects of glioma. Immunofluorescence analyses of Kv1.3 expression (red) on GFAP-positive cells (green) in coronal brain slices of GL261-bearing mice in peritumoral region; in the magnification (right) arrows indicate co-expression of Kv1.3 and GFAP in the same cell. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-25940-5) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: GFAP Antibody [NB300-141] - Mouse brain section, 20x magnification. Antibody at 1:1000. Detection with Polymer-HRP. IHC image submitted by a verified customer review.

Western Blot: GFAP Antibody [NB300-141] - Analysis of Rat brain lysate. Antibody at 1:5000. Specific band running with an apparent SDS-PAGE molecular weight of ~50 kDa corresponds to rodent GFAP was observed.

Western Blot: GFAP Antibody [NB300-141] - Analysis of GFAP expression in whole rat cerebellum homogenate.

Western Blot: GFAP Antibody [NB300-141] - Analysis of different tissue lysates using rabbit polyclonal antibody to GFAP, NB300-141, dilution 1:5000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord. Strong band at about 50 kDa corresponds to the major isotype of the GFAP protein. Smaller isotypes and proteolytic fragments of GFAP are also detected on the blot.

Immunocytochemistry/Immunofluorescence: GFAP Antibody [NB300-141] - Analysis of mixed neuron-glial cultures using GFAP antibody NB300-141 (red) and Vimentin antibody NB300-223 (green). The fibroblastic cells contain only Vimentin and so are green. The astrocytes contain either Vimentin and GFAP (appearing golden) or predominantly GFAP (appearing red). Blue is nuclear DNA stain.

Immunocytochemistry/Immunofluorescence: GFAP Antibody [NB300-141] - Cultured Rat hippocampal neurons. ICC/IF image submitted by a verified customer review.

Immunohistochemistry: GFAP Antibody [NB300-141] - Xenografted mouse brain section: astocyte and human nuclei. ICC/IF image submitted by a verified customer review.

Immunohistochemistry-Frozen: GFAP Antibody [NB300-141] - Imaging of mouse brain (cortex), 20x magnification. IHC image submitted by a verified customer review.

EGFP+ neurons are positive for RABV antigen.Brains were collected from Cre reporter mice fifteen days post-infection, cryosectioned, and EGFP+ regions compared to cell-specific labeling, A) NeuN (blue, neuronal nuclei antibody, 20× fluorescence imaging), B) GFAP (blue, astrocyte antibody, 40× confocal imaging), or C) RABV P antigen (purple) and DAPI nuclear stain (blue, 63× confocal imaging). White arrows in (C) indicate regions positive for RABV P.

Immunocytochemistry/ Immunofluorescence: GFAP Antibody [NB300-141] - EGFP+ neurons are positive for RABV antigen.Brains were collected from Cre reporter mice fifteen days post-infection, cryosectioned, & EGFP+ regions compared to cell-specific labeling, A) NeuN (blue, neuronal nuclei antibody, 20× fluorescence imaging), B) GFAP (blue, astrocyte antibody, 40× confocal imaging), or C) RABV P antigen (purple) & DAPI nuclear stain (blue, 63× confocal imaging). White arrows in (C) indicate regions positive for RABV P. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1002971), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GFAP Antibody [NB300-141] - Effects of the repeated administration of NVP CXCR2 20 (NVP; 10 μg/5 μl; i.t.; 16 h & 1 h before CCI & then once a day for 7 days) on the protein levels of CXCR2, IBA1, GFAP, CXCL1, CXCL2, & CXCL3 proteins (A–I) in the spinal cord (A–F) & DRG (G–I) on the 7th day after CCI in rats. The data are presented as the mean fold changes relative to the control ± SEM (5–6 samples per group). Intergroup differences were analyzed using ANOVA with Bonferroni's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001 indicate differences vs. naive rats. #p < 0.05, indicate differences between V-treated & NVP-treated rats. CCI, chronic constriction injury; N, naive; V, vehicle; NVP, NVP CXCR2 20. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31616413), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFAP Antibody [NB300-141] - Increased astrocyte activation in IL-1 beta -stimulated two-week old offspring is reduced by maternal LB supplementation. Representative images of fluorescence microscopy of claudin-5+ (location of the brain capillaries, red), GFAP+ astrocyte (green), & DAPI (nuclei, blue). Three to five sections per mouse were examined & at least three mice were examined in each group. Stronger than control SPF GFAP staining was observed around the blood vessel (see arrow) after IL-1 beta insult. Maternal supplemented group (LB) with or without postnatal insult had GFAP levels similar to the control group indicating that LB supplementation prevented astrocyte activation around the BBB endothelium. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32424168), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFAP Antibody [NB300-141] - Glial cells density is differently modulated during AD progression.a Retinal slices were immunolabeled with anti-GFAP antibody (green) & Hoechst for nuclei visualization (blue) at different ages of 3xTg-AD & non-Tg mice & density of GFAP signal was quantified as shown in b (**p < 0.01 pre vs early; n = 16 fields/four slices for each condition; two-way ANOVA, Holm-Sidak; ##p < 0.01 for comparison with age-matched non-Tg mice, two-way ANOVA, Holm-Sidak). c Representative multiarea image of retinal slice immunolabeled with anti-Iba1 antibody (green) & Hoechst for nuclei visualization (blue); density of Iba1+ cells was quantified as shown in d (**p < 0.01 vs 3xTg-AD pre; ##p < 0.01 vs age-matched non-Tg mice, n = 16 fields/four slices for each condition, two-way ANOVA, Holm-Sidak method for multiple comparison) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29880901), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ch, Eq, Gp, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ch, Eq, Gp, Rb

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NB600-633

IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

(4 Reviews)

(116 Publications)

0.1 mg

NB600-633

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody [NB600-633] - Analysis of: Human IL1beta antibody Secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT Localization: cytoplasm Staining: Close up of medullary lymph node: positive staining in the cytoplasm of circulating macrophages. Neg Ctr (far right) normal rabbit IgG with pH 6.2 40X

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Analysis of IL-1 beta in Rhesus fetal membrane using anti-IL-1 beta antibody. Image from verified customer review.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Antibody was used at a 1:200 dilution incubated 1h at room temperature to detect dog IL-1b by Western blot. Lane 1 and 2 were loaded with 2.5 ug and 500 ng of dog IL-1b respectively. The molecular weight of the detected band is estimated by comparison to molecular weight markers (not shown). Detection occurred using a 1:3,000 dilution of IRDye (TM) 800 conjugated Donkey anti-Rabbit IgG for 1h at room temperature.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - pro-IL-1beta in glioblastoma cells. Image from verified customer review.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Lane 1: Human IL-1Beta. Load: 5 ng per lane. Primary antibody: Human IL-1Beta antibody at 1:2,000 for overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT. Predicted/Observed size: 17 kDa, 17 kDa for Human IL-1Beta. Other band(s): Unspecific band at 35 kDa.

Western Blot of Rabbit anti-Human IL-1beta antibody. Lane 1: Human IL-1beta. Load: 5 ng per lane. Primary antibody: Human IL-1beta antibody at 1:2,000 for overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting () for 30 min at RT. Predicted/Observed size: 17 kDa, 17 kDa for Human IL-1beta. Other band(s): Unspecific band at ~35 kDa.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Methylene blue inhibits inflammasome formation in cultured microglia. (A) Expression of Pro-Caspase-1, Caspase-1 p20, pro-IL-1 beta & mature IL-1 beta in microglia with or without stimulation in the presence or absence of methylene blue. Left: representative Western blot images. Right: statistics. N = 5 per group. The (B) expression of NLRP3, NLRC4 & Aim2 in microglia. Left: representative Western blot images. Right: statistics. N = 4 per group. (C) Co-immunoprecipitation assay showing the binding of ASC to NLRP3 or NLRC4. Left: representative Western blot images. Right: statistics. Un: no stimulation. LPS+ATP: stimulation with LPS followed by ATP treatment. V: vehicle; M: 500 nM methylene blue. IP: immunoprecipitation with the antibody against indicated protein. IB: detection of indicated protein. To analyze the relative expression of target proteins, the band intensities of target proteins were normalized to corresponding band intensities of alpha -Tubulin, followed by calculating the expression in other groups relative to “Un (V)” group. N = 4 per group. *p < 0.05; **p < 0.01; ***p < 0.001 in comparison with unstimulated cells of the vehicle group. #p < 0.05; ##p < 0.01 in comparison with stimulated cells of the vehicle group. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fncel.2017.00391/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Rabbit Polyclonal IL-1 beta/IL-1F2 Antibody [NB600-633] - IL-1 beta/IL-1F2 Western blot of liver homogenate from control (1) and alcohol-fed mice (2). Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Po, Ca, Pm, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Pm, SyHa

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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HAF017

Rabbit anti-Goat IgG Secondary Antibody [HRP (Horseradish Peroxidase)]

(1 Review)

(123 Publications)

1 mL

HAF017

Western blot shows detection of Goat Anti-Human/Mouse/Rat Syntaxin 1A Polyclonal Antibody (Catalog # AF7237) using a 1:1000 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Evaluation of ZL-2201 antiproliferative activity in vitro. A, Chemical structure of ZL-2201, a potent and selective DNA-PK inhibitor. B, Concentration-dependent response to ZL-2201 in M059J and M059K glioblastoma cancer cells measured by CTG after 6-day of treatment. The graph represents the average inhibitory (IC50) values (n = 3). C, Concentration-dependent response to ZL-2201 in CRISPR KO of ATM in A549 and FaDu cancer cells measured by CTG after 6 days of treatment. The graph represents the average inhibitory (IC50) values (n = 2–5). D, IC50 responses to ZL-2201 in ATM mutant cell lines (black bars) versus ATM WT A549 cell line (gray bar). Cell growth was measured by CTG after 6 days of treatment. The graph represents the average inhibitory (IC50) values (n = 2–6). NCI-H1703 (E) and A549 (F) cancer cells were treated with bleomycin (10 μmol/L) for 2 hours followed by the addition of increasing concentration of ZL-2201 for 2 hours. Whole-cell lysates were harvested, and concentration-dependent inhibition of DNA-PK protein were analyzed by Simple Western. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/37663435), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Gt

Species Glossary

Reactivity: Gt

Applications:

Applications:

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

HRP (Horseradish Peroxidase)

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF017

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	R-401-C-ABS

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NB600-363

HA Tag Antibody - BSA Free

(3 Reviews)

(111 Publications)

0.1 ml

NB600-363

Western Blot: HA Tag Antibody [NB600-363] - Wnt/beta-catenin signaling regulates HySp5 expression. Immunoprecipitation (IP) of HA-tagged HySp5-420 expressed in HEK293T cells together or not with hudeltabeta-Catenin (upper) or huTCF1 (lower). IP was performed with an anti-HA antibody and Co-IP products were detected with the anti-beta-catenin or anti-TCF1 antibodies. Same results were obtained in two independent experiments. Each data point represents one biological independent experiment. Statistical p values: *p <= 0.05, **p <= 0.01, ***p <= 0.001, ****p <= 0.0001 (unpaired t test). Error bars indicate SD. PP primer pair, RLA relative luciferase activity Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-08242-2), licensed under a CC-BY license.

Western Blot: HA Tag Antibody [NB600-363] - ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors and pEGFP, cell lysates were probed with antibodies against the HA tag and EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, and 1200 ng. NT, non-transfected cells. Selection-independent generation of gene knockout mouse embryonic stem cells using zinc-finger nucleases. PLoS One (2011)

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - HA-tagged proteins were detected in immersion fixed HEK293 human embryonic kidney cell line transfected with HA-tagged MGAT2 using 1 ug/mL goat anti-HA Tag polyclonal (NB600-362, Novus Biologicals), 1 ug/mL rabbit anti-HA Tag polyclonal (NB600-363, Novus Biologicals). Cells were stained using the appropriate secondary antibody, donkey anti-mouse IgG-NL557 (NL007) and counterstained with DAPI (blue).

Western Blot: HA Tag Antibody [NB600-363] - Gene knockout in U2OS.693 cells.(A) Schematic of ZFN-mediated knockout. A ZFN pair (ZFNR & ZFNL) designed to target position 502 in the EGFP gene (E502) creates a DNA double-strand break that is sealed by the error-prone non-homologous end-joining (NHEJ) pathway & hence leads to disruption of the coding sequence. (B) Dose-dependent gene disruption in U2OS.693 cells. U2OS.693 cells that stably express a destabilized EGFP were transfected with increasing amounts of E502-specific ZFN expression vectors (75–1200 ng). The percentage of EGFP-negative cells was determined 6 days post-transfection by flow cytometry (n = 3; indicated is average & standard deviation). E502-WT, ZFN with wild-type FokI domain; E502-OH, ZFN with obligate heterodimeric FokI domain. (C) ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors & pEGFP, cell lysates were probed with antibodies against the HA tag & EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, & 1200 ng. NT, non-transfected cells. (D) Kinetics of EGFP knockout. The graph displays the percentage of EGFP-negative cells (see B) from day 1 to day 6 post-transfection for two vector amounts (n = 3; indicated is average & standard deviation). WT, E502-WT; OH, E502-OH. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22194948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

ET

Species Glossary

Reactivity: ET

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-363AF350
Alexa Fluor 405	NB600-363AF405-0.1ml
Alexa Fluor 488	NB600-363AF488-0.1ml
Alexa Fluor 532	NB600-363AF532
Alexa Fluor 594	NB600-363AF594
Alexa Fluor 647	NB600-363AF647-0.1ml
Alexa Fluor 700	NB600-363AF700-0.1ml
Alexa Fluor 750	NB600-363AF750
Allophycocyanin	NB600-363APC
Biotin	NB600-363B
CoraFluor 1	NB600-363CL1
DyLight 350	NB600-363UV-0.1ml
DyLight 405	NB600-363V-0.1ml
DyLight 488	NB600-363G-0.1ml
DyLight 550	NB600-363R-0.1ml
DyLight 594	NB600-363DL594
DyLight 650	NB600-363C-0.1ml
DyLight 680	NB600-363FR-0.1ml
DyLight 755	NB600-363IR-0.1ml
FITC	NB600-363F
HRP	NB600-363H-0.1ml
Janelia Fluor 525	NB600-363JF525
Janelia Fluor 549	NB600-363JF549
Janelia Fluor 646	NB600-363JF646
Janelia Fluor 669	NB600-363JF669
PE	NB600-363PE
PerCP	NB600-363PCP
mFluor Violet 450 SE	NB600-363MFV450
mFluor Violet 500 SE	NB600-363MFV500
mFluor Violet 610 SE	NB600-363MFV610

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2275-PC-100

GAPDH Antibody [Unconjugated]

(111 Publications)

100 uL

2275-PC-100

Western blot shows lysates of U2OS human osteosarcoma cell line, A20 mouse B cell lymphoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat GAPDH Polyclonal Antibody (Catalog # 2275-PC) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for GAPDH at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western shows lysates of Exosome Standards (LNCaP) (NBP3-11687), Exosome Standards (HT‑29) (NBP3-11685) and Jurkat human acute T cell leukemia cell line, loaded at 0.5 mg/ml. A specific band was detected for GAPDH at approximately 41 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # 2275-PC-100). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, C2C12 mouse myoblast cell line, and C6 rat glioma cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH at approximately 43 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # 2275-PC-100) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Higher levels of CD4 are found in macrophages expressing LCK.A) Schematic diagram of self-inactivating lentiviral construct used to express wild-type Lck and puroR (LCKWT P). A similar construct was used to express a kinase inactive form of Lck and puroR (LCKINACTIVE P) or to express puroR only (‘control P’). B) Detection of protein expression of LCK and CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-LCK and anti-CD4 antibodies. The loading control GAPDH was detected using anti-GAPDH antibody. C) Protein levels were measured with Odyssey software (Li-COR) and CD4 expression was normalised to GAPDH expression. Symbols represent normalised CD4 expression, relative to the PSC-macrophages control group, of two independent experiments. D) Detection of surface CD4 and total LCK expression. Representative two-colour immunofluorescence (dot plot) analysis is shown. Gates were determined by using the two relevant isotype control antibodies. Quantification of total LCK expression (E) and surface CD4 expression (F), expressed as the ratio of the geometric mean fluorescence intensity (MFI) over the isotype control ±SEM of independent experiments (n = 7). G) Detergent resistance of CD4, expressed as mean Flow Cytometric Detergent Resistance (FCDR) index of CD4 (n = 4) in PSC-macrophages, calculated as described in materials and methods. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.

CD4 knock-down in genetically modified stem cell-derived macrophages. C) Detection of protein expression of CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-CD4 antibodies. GAPDH, a loading control, was detected using anti-GAPDH antibody. Representative blot is shown. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NBP1-49461

Stathmin-2/STMN2 Antibody - BSA Free

(2 Reviews)

(102 Publications)

0.1 mg

NBP1-49461

<b>Genetic Strategies Validation. </b>Staining of STMN2 in primary mouse dorsal root ganglia (DRG) neurons Image shows the expected staining of endogenous STMN2 in axons and growth cones. Tubulin is a marker of axons. NBP1-49461 was used at a dilution of 1:4000. Image courtesy of Dr. Jung Eun Shin.

Neuronal deletion of Tsc2 improves axon regeneration within the first 2 d following nerve injury. SCG10 immunostaining of sciatic nerves 1 d after crush injury in control and Tsc2 cKO mice with (conditioned) or without (sham) a conditioning injury. Red dotted line denotes the crush site while red arrowheads point to the three longest axons. Scale bar: 500 i1/4m. Image collected and cropped by Citeab from the following publication (Nociceptor Deletion of Tsc2 Enhances Axon Regeneration by Inducing a Conditioning Injury Response in Dorsal Root Ganglia. Eneuro (2019) licensed under a CC-BY license.

<b>Expression of Tagged Proteins Validation. </b>Staining of STMN2 in HEK293T cells transfected with GFP or a GFP-STMN2 fusion. NBP1-49461 was used at a dilution of 1:1000 (A, B) and 1:3000 (C). Image courtesy of Dr. Jung Eun Shin.

<b>Expression of Tagged Proteins Validation. </b>Analysis of STMN2 in HEK293T cells transfected with GFP or a GFP-STMN2 fusion. NBP1-49461 was used at a dilution of 1:4000. Image courtesy of Dr. Jung Eun Shin.

Reactivity:

Hu, Mu, Rt, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Bv

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-49461AF350
Alexa Fluor 405	NBP1-49461AF405
Alexa Fluor 488	NBP1-49461AF488
Alexa Fluor 532	NBP1-49461AF532
Alexa Fluor 594	NBP1-49461AF594
Alexa Fluor 647	NBP1-49461AF647
Alexa Fluor 700	NBP1-49461AF700
Alexa Fluor 750	NBP1-49461AF750
Allophycocyanin	NBP1-49461APC
Biotin	NBP1-49461B
CoraFluor 1	NBP1-49461CL1
DyLight 350	NBP1-49461UV
DyLight 405	NBP1-49461V
DyLight 488	NBP1-49461G
DyLight 550	NBP1-49461R
DyLight 594	NBP1-49461DL594
DyLight 650	NBP1-49461C
DyLight 680	NBP1-49461FR
DyLight 755	NBP1-49461IR
FITC	NBP1-49461F
HRP	NBP1-49461H
Janelia Fluor 525	NBP1-49461JF525
Janelia Fluor 549	NBP1-49461JF549
Janelia Fluor 646	NBP1-49461JF646
Janelia Fluor 669	NBP1-49461JF669
PE	NBP1-49461PE
PerCP	NBP1-49461PCP
mFluor Violet 450 SE	NBP1-49461MFV450
mFluor Violet 500 SE	NBP1-49461MFV500
mFluor Violet 610 SE	NBP1-49461MFV610

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NBP1-05987

ZEB1 Antibody

(6 Reviews)

(102 Publications)

0.1 ml

NBP1-05987

Western Blot: ZEB1 Antibody [NBP1-05987] - Analysis of ZEB1 in transfected and native mouse cell lysate. Image courtesy of anonymous customer review.

Immunohistochemistry-Paraffin: ZEB1 Antibody [NBP1-05987] - Analysis of ZEB1 in human liver and spleen tissues. Image courtesy of anonymous customer review.

<b>Independent Antibodies Validation. </b>Immunoprecipitation: ZEB1 Antibody [NBP1-05987] - Detection of human ZEB1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-ZEB1 antibody NBP1-05987 used for IP at 3 ug per reaction. ZEB1 was also immunoprecipitated by another rabbit anti-ZEB1 antibody. For blotting immunoprecipitated ZEB1, NBP1-05987 was used at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

Immunohistochemical staining results of EMT regulators in HG‐PanIN: SNAIL, SLUG, TWIST1, and ZEB1. We categorized and defined three grades: grade 0, <10% positive staining; grade 1, 10%‐50% positive; grade 2, 50%< positive

Western Blot: ZEB1 Antibody [NBP1-05987] - Lin28a & ETM-related protein expression changed after the knockdown of Zeb1 in mESCs, mEB, GC1, & P19. Western blotting demonstrated that E-cadherin protein expression increased & that Lin28a & Vimentin protein expression decreased in Zeb1-siRNA transfected mESC (panel a), mEB (panel b), GC1 (panel c), & P19 (panel d) cells. Each experiment was performed in triplicate. Abbreviations: NC, negative control; siRNA, small interfering RNA; & WT, wild-type. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35611144), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: ZEB1 Antibody [NBP1-05987] - Immunohistochemical staining results of EMT regulators in HG‐PanIN: SNAIL, SLUG, TWIST1, & ZEB1. We categorized & defined three grades: grade 0, <10% positive staining; grade 1, 10%‐50% positive; grade 2, 50%< positive Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30791220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: ZEB1 Antibody [NBP1-05987] - Location of Zeb1 proteins in mEC lines (mESC, mEB, GC1, & P19). Immunofluorescence analysis was used to determine localization of Zeb1 in mESC, mEB, GC1, & P19 using a Zeb1 antibody (red). Cell nuclei were labeled with Hoechst 33342 (blue). The scale for the measurement bar is 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35611144), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - Zeb1 expression in mESCs, mEB, GC1, & P19. (a) Zeb1 mRNA expression detected by real-time qPCR in mESC, mEB, GC1, & P19 (P < 0.001, data presented as mean value ± SD). (b) Zeb1 protein expression examined by western blot in mESC, mEB, GC1, & P19. Each experiment was performed in triplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35611144), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: ZEB1 Antibody [NBP1-05987] - Correlation of the expression of ZEB1 & IL‐6 & other cytokines in various cancers. (A) Correlation analysis of the expression of ZEB1 & cytokines in breast cancer cell lines using data obtained from the Cancer Cell Line Encyclopedia (CCLE). Breast cancer cell lines were selected for the analysis, & each dot represents a cell line. Affymetrix microarray probe IDs for IL6 & IL8 are shown after the gene symbols on the y‐axis. RMA, robust multiarray average. (B) Representative results obtained from the tissue array analysis in Fig. S3. The top panel shows a case that was positive for both nuclear ZEB1 staining (green) & whole‐cell IL‐6 staining (red); scores = 4. The bottom panel shows a case that was negative for both ZEB1 & IL‐6; scores = 1. The samples were counterstained with DAPI to show cell density in the spot. Original magnification: 20 ×. (C) Kaplan–Meier survival curves of breast & lung cancer patients obtained from a public meta‐analysis database & Kaplan–Meier plotter (Gyorffy et al., 2010, 2013). The probability of overall survival of patients as split by median IL6 & IL8 expression is shown. Red: IL6 & IL8 high expression group; black: IL6 & IL8 low expression group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28618162), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - SU5402, FGFR1 inhibitor, affects the EMT transcription factors.(A, B) ERK1/2 phosphorylation (p-ERK1/2) determined by immunoblotting in HSC-4 & OTC-04 treated for 30 min w/ 30 ng/ml FGF2 or 30 ng/ml FGF7 in presence of 10% FBS (A) & in TSU & HOC313 cells treated for 1 h w/ 30 ng/ml FGF2 or 30 μM SU5402 in absence of FBS (B). F2, FGF2; F7, FGF7; SU, SU5402. (C) We have previously reported that, after treatment w/ TGF-beta , NMuMG cells underwent EMT w/ the IIIc-isoform of FGFR1[17]. After NMuMG cells pretreated w/ TGF-beta transfected w/ mouse Fgfr1 siRNA or treated w/ SU5402, the cells further incubated in culture medium (CM) from TSU cells. SU, SU5402; siFR1, siRNA against mouse Fgfr1. (D)FGF2 mRNA levels determined by RT-qPCR analyses. The ratio of FGF2 mRNA to GAPDH mRNA in HSC-4 cells indicated as “1”. Each value represents the mean ± SD of triplicate determinations from a representative experiment. Similar results obtained in at least three independent experiments. (E) ERK1/2 phosphorylation (p-ERK1/2) in TSU & HOC313 cells monitored in presence of indicated concentration of SU5402 for 1 h under serum-free culture conditions, followed by immunoblot analysis. (F, G) Expression of indicated genes in TSU cells under serum-free culture conditions determined by RT-qPCR (F) & immunoblot (G) analyses, following treatment w/ 10 μM SU5402. Each value represents the mean ± SD of triplicate determinations from a representative experiment. Similar results obtained from at least three independent experiments. p values determined by Student’s t-test. **p < 0.01. (H) TSU cells treated w/ 10 μM U0126 in absence of FBS subjected to immunoblotting w/ the indicated antibodies. alpha -tubulin used as a loading control (A, B, C, E, G, & H). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31682640), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - Vimentin & ZEB1 protein levels are regulated by OVOL1 & OVOL2 in a human SCC cell line. (A) Relative protein expression levels of OVOL1 & OVOL2 in A431 cells treated with control siRNA, OVOL1 siRNA, or OVOL2 siRNA for 48 h. (Left) Representative blot images & (right) relative expression levels calculated from three independent experiments. (B) Relative protein expression levels of vimentin, ZEB1, & E-cadherin in A431 cells treated with control siRNA, OVOL1 siRNA, or OVOL2 siRNA for 72 h. (Upper) representative blot images & (lower) relative protein expression levels calculated from three independent experiments. Mann–Whitney U-test; error bars represent mean ± standard deviation. Protein expressions are relative to those of beta -actin as a reference. The control siRNA value was set to 1. p-values < 0.05 were assumed to indicate a statistically significant difference; * p < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32106476), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - PDGFR-alpha / beta promotes sarcoma CSC migration/invasion & anchorage-independent growth.a Western blot analysis of cell adhesion protein N-cadherin & EMT transcription factors Snail, Slug, & Zeb1 in human sarcoma cell lines grown as monolayers & as spheroids. beta -actin was used as the loading control. b Representative images under light microscopy of migration & invasion assays of human sarcoma cell lines grown as monolayers or as spheroids for 24–48 h. Graphs display the number of migrated or invasive cells per field. c Representative light microscopy images of forrmed colonies in soft agar assay of human sarcoma cell lines grown as monolayers or spheroids for 14–20 days. Graph displays the number colonies per field. d Representative images under light microscopy of migration & invasion assays of human sarcoma cell lines grown as monolayers or as spheroids. Spheroid cells were treated with imatinib or DMSO. e Western blot analysis of N-cadherin & EMT-regulating transcription factors in human sarcoma cell lines grown as spheroids & treated with imatinib or DMSO. Experiments in a & e were performed three times with similar results. Bars represent standard deviation. *p < 0.05 compared to Monolayers Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41389-018-0059-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - FGFR1 siRNAs attenuate the malignant phenotypes of cancer cells.(A) mRNA from TSU cells transfected with siRNAs against FGFR1 (siFGFR1) were subjected to conventional RT-PCR to determine the levels of endogenous FGFR1. (B, C, D) After transfection with siFGFR1 in TSU cells, phosphorylation of ERK1/2 (p-ERK1/2) (B), ZEB1 levels (C), & cell morphology (D) were determined. (E) TSU cells transfected with siFGFR1 were subjected to immunofluorescence analyses. Low magnification, 40×; high magnification, 80×. (F, G, H, I) After either treatment with SU5402 or transfection with siFGFR1 in TSU & HOC313 cells, the number of cells & invasive properties were determined under serum-free culture conditions. (J & K) After transfection with siFGFR1c in TSU & HOC313 cells, invasive & migratory properties were determined under serum-free culture conditions in TSU & HOC313 cells, respectively. siNC, non-specific control siRNA. Each value represents the mean ± SD of triplicate determinations from a representative experiment. Similar results were obtained from at least three independent experiments. p values were determined by Student’s t-test. *p < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31682640), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - Nav1.4 protein levels are decreased in muscles from mouse models of SMA. (A) Immunoblot analysis using muscle lysate from P2, P5, P9, & P21 wild type mice. Nav1.4 protein levels increase during postnatal muscle development & form the predominant sodium channel expressed in mature skeletal muscle. GAPDH served as a loading control (N = 3). (B) Representative immunoblot with quantification, showing a decrease in levels of sodium channel Nav1.4 & Nav1.5 in P5 Smn-/-;SMN2 hindlimb muscle compared with controls (N = 3). (C) Quantification of immunoblot analyses in P21 Smn2B/- & control hindlimb muscles revealed a decrease in Nav1.4 levels. Early in postnatal muscle development, the Nav1.5 sodium channel isoform is the most predominant. In P21 Smn2B/- mice, the protein levels of Nav1.5 are higher than in controls (N = 3). (D) The protein level of the Nav1.4 positive regulator, NF1, is not altered in muscles from P21 Smn2B/- mice. Similarly, no change was detected in the protein levels of the Nav1.4 repressor ZEB. (E) Expression of sodium channel Nav1.4 in control sham & denervated samples 1 & 7 days post-denervation was assessed by immunoblot (N = 3). A decrease in the levels of Nav1.4 in muscle was noted at 7 days post-denervation. *, P < 0.05; **, P < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24119341), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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Validated by:

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NB100-137

EGLN1/PHD2 Antibody - BSA Free

(12 Reviews)

(96 Publications)

0.1 mg

NB100-137

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Analysis of PHD2 expression in human prostate cancer cell lines. Western blot image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-137] - Staining of endogenous PHD2 in U2OS cells. ICC/IF image submitted by a verified customer review.

<b>Genetic Strategies Validation. </b>Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Detection of PHD2 in human glioblastoma cell lysates. Image submitted by a verified customer review.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Detection of human PHD2 by Western blot. Recombinant epitope-tagged PHD1, PHD2 or PHD3 (10 ng/lane) or whole cell lysate from HeLa cells. EGLN1/PHD2 antibody used at 1 ug/mL. Detection by chemiluminescence.

Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-137] - EGLN1/PHD2 and EGFR expression levels positively correlate in breast cancer. Processed tissue microarrays of breast cancer biopsies from 313 patients were stained with EGLN1/PHD2 and EGFR antibodies. Four representative immunohistochemistries of human breast cancer with low and high expression of EGLN1/PHD2 and EGFR are shown. Magnification 10x. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/article/14241/text/) licensed under a CC-BY license.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Exogenous PHD2 has no impact on the gene expression of HIF-1alpha and HIF-2alpha. U87MG cells were transfected with empty vector (pcDNA) or PHD2 plasmid. Twenty-four hours after transfection, PHD2 was detected by immunoblotting. beta-Actin was used as loading control. The results are representative for three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014295), licensed under a CC-BY license.

Flow Cytometry: EGLN1/PHD2 Antibody [NB100-137] - An intracellular stain was performed on Jurkat cells with NB100-137AF647 (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.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Analysis of PHD2 in normoxia (lane 1) or hypoxia (lane 2) treated SK-N-BE(2) cells using anti-EGLN1/PHD2 antibody. Western blot image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-137] - Staining of PHD2 in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-137] - Staining of lung vascular endothelium with EGLN1/PHD2 antibody. Image at 40X.

Flow Cytometry: EGLN1/PHD2 Antibody [NB100-137] - Detection of PHD2 in Jurkat cells. One million Jurkat cells were fixed, permeabilized, and stained with 3.0 ug/mL anti-EGLN1/PHD2 antibody in a 150 uL reaction. Isotype control (black), anti-MLL1 (red).

PHD2 and EGFR expression levels positively correlate in breast cancerProcessed tissue microarrays of breast cancer biopsies from 313 patients were stained with PHD2 and EGFR antibodies (cf Materials and methods). Four representative immunohistochemistries of human breast cancer with low and high expression of PHD2 and EGFR are shown. Magnification 10x.

Initial characterization of the OS-9 protein.(A) OS-9 expression in various human cell lines. Equal protein amounts of total cell lysates were used for SDS-PAGE & subsequent Western blotting. For each cell line, two independent samples are shown. Endogenous OS-9 was detected with a polyclonal antibody raised against a peptide corresponding to amino acids 600–667 of isoform 1 of OS-9. (B) Protein stability assay of endogenous OS-9. U2OS cells were treated with the translational inhibitor cycloheximide (100 µM). At indicated time points, whole cell lysates were analysed by immunoblotting. (C) Effect of hypoxia on OS-9 expression. For hypoxia, UT-7 cells were exposed to 1% O2 for 24 h prior to Western blot analysis. To determine any influence of HIF-1 alpha on OS-9 expression under normoxia, cells were incubated with the prolyl hydroxylase inhibitor DMOG (0.5 mM) for 24 h. (D) Protein interaction between OS-9 & PHD2 in vitro. For co-immunoprecipitation, U2OS cells were transiently co-transfected with the plasmids pOS-9-V5 & pPHD2-His, lysed in NP40 buffer, & subjected to immunoisolation with anti-V5 antibody recognizing OS-9 by its V5-tag. OS-9 & its associated proteins were separated by SDS-PAGE & analyzed by Western blot (lane 2). As controls, samples of untransfected (lane 1) cells or cells transfected with a single plasmid (lanes 3–4) were loaded. Representative Western blots are shown for each subfigure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Cellular localization of OS-9 & PHD2.(A) A lectin gel-shift assay was conducted to test for glycosylated proteins. Total cell lysates of U2OS cells were incubated in the presence or absence of the endoglycosidases EndoH & PNGaseF for 6 h at 37°C. Digest products were separated on a reducing 10% SDS-PAGE gel which contained concanavalin A co-polymerized in the top layer of the separating gel to retard mobility of glycosylated proteins [56]. Glycosylated OS-9 is indicated as ‘g’, deglycosylated OS-9 as ‘d’. (B) Detection of OS-9 & PHD2 in the nuclear fraction. HEK293 cells with & without transfection of the plasmid pcDNA3-OS-9 were separated into nuclear fraction (N) & postnuclear supernatant (PS), the latter containing cytoplasm & organelles. Western blot analysis included BiP, GAPDH & lamin A as typical marker proteins for the ER, the cytoplasm & the nucleus, respectively. (C) Detection of OS-9 & PHD2 in the cytoplasm. HEK293 cells were co-transfected with pcDNA3-OS-9 & pPHD2-V5. For hypoxia, cells were exposed to 3% O2 for 4 h. Cells were treated with 50 µg/ml digitonin & centrifuged to obtain a cytoplasmic (C) & an organelle fraction (O) & subjected to immunoblotting. (D) Isolation of cellular endomembranes. HEK293 cells were lysed mechanically by several passages through a 30½G needle. The postnuclear supernatant was processed further by ultra-centrifugation to separate the organelles (O) from the cytosol (C). High salt treatment (1 M KCl) of the organelle fraction produced a wash fraction (W) that contained dissociated peripheral membrane proteins. For immunoblot analysis of subcellular fractionations, cell aliquots were normalized for cell number prior to loading (B–D). Representative Western blots are shown for each subfigure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Representative picture of western blot in histopathologically unchanged tissue (N) & primary cancerous tissue (C) from patients with CRC. Immunodetection of bands was performed with Rp anti- PHD1, - PHD2, - PHD3 & - FIH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The membrane was stripped & incubated with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. Bands were revealed using SuperSignal West Femto Chemiluminescent Substrate, Thermo Fisher Scientific (Rockford, IL) & Biospectrum® Imaging System 500, UVP Ltd. (Upland, CA). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - OS-9 shows no effect on regulation of HIF-1 alpha .Total cell lysates were used for SDS-PAGE & subsequent Western blotting. To generate nearly anoxic conditions, cells were exposed to an oxygen consuming chemical system or to 1% or 3% O2 for 4 h to generate hypoxia. (A) U2OS, HeLa & Hep3B cells were transiently transfected with the plasmid pOS-9-V5 48 h prior to the experiment. Lamin A & actin were used as loading controls. (B) U2OS cells were subjected to ER stress by incubation either with tunicamycin (1 µg/ml) or thapsigargin (0.5 µg/ml) for 20 h. To detect HIF-1 alpha under normoxia, cells were treated with DMOG (1 mM) for 4 h. A sample of DMSO-only treated cells was loaded to exclude unspecific side effects of the solvent. (C) U2OS cells were transduced with lentiviral construct pLKO.1-shRNA-OS-9 (shOS-9) mediating a stable knockdown of OS-9 expression. Control cells (c) were transduced with plasmid pLKO.1-puro. Representative Western blots are shown for each subfigure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Oxygen chemotaxis is independent of the PHD/HIF pathway. a–dHIF1A, HIF2A, HIF1A+HIF2A & PHD2 CRISPR/Cas9 KO clones characterisation, respectively, regarding O2-directed migration. Left panels: immunoblot validation of HIF1A, HIF2A, HIF1A+HIF2A & PHD2 KO clones. To blot HIFs factors (a, b, c), cells were first pre-treated for 5 h with CoCl2 300 µM before protein extraction, a condition that promotes HIF factors accumulation (cf. Supplementary Fig. 8f). Middle panels: cell trajectories under confinement. Red dashed lines indicate the border of the cell cluster at 0 h. Right panels: relative distribution of MCF10A KO clones versus wt cells at the edge of the cluster at 48 h. These experiments demonstrate that HIF factors deletion does not prevent aerotaxis. e, f Tracks & redistribution of wt & PHD2 KO clone silenced for PHD3 (siPHD3) or not (siCTR). g Tracks & redistribution of MCF10A cells treated with DMOG (50 µM) or CoCl2 (50 µM) to inhibit PHDs, or with vehicle only (DMSO). These experiments demonstrate that PHDs do not participate in O2-sensing during aerotaxis. Confinement was applied for 48 h (a–g). Scale bars, 500 µm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30382089), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - PHD3 expression correlates with a mesenchymal-like morphology in pancreatic ductal adenocarcinoma cell lines.NHF-1 (Fibroblast) MiaPaca2, Panc1, CAPAN1 & BxPC3 cells were harvested for RNA & protein following 24 hours exposure to normoxia (21% O2) or hypoxia (1% O2). (A) Phase-contrast images at 10× magnification were taken of MiaPaca2 (mesenchymal-like) morphology & BxPC3 cells (differentiated, epithelial morphology) under normoxic conditions. (B) PHD3 mRNA expression was determined by qRT-PCR & graphed relative to BxPC3 in normoxia. All samples were normalized to 18S rRNA & graphed as expression relative to BxPC3-Vec Normoxia (lane 1). n = 3, error bars = 1 S.D. (C) Whole cell lysate was resolved by SDS-PAGE & blotted for beta -tubulin PHD3 & PHD2. N = normoxia, H = hypoxia (1% O2). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0083021), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Iron supplementation restores HIF-1 alpha levels to normal following ATP6V0E1 inhibition in HeLa cells. (A) Schematic diagram of the multimeric V-ATPase complex. (B) Chemical inhibition of V-ATPase by 10 nM BafA treatment for 24 h, increased HIF-1 alpha levels in HIF alpha -GFPODD reporter cells. Treatment with 100 μM Fe (III) citrate significantly reduced the elevated HIF-1 alpha levels associated with loss of ATP6V0E1 (1 × 106 cells per sample harvested & analyzed; N = 2). (C) Knock-down of ATP6V0E1 subunit with three different CRISPR-Cas9 guides resulted in significant upregulation of HIF-1 alpha levels in HIF alpha -GFPODD reporter cells. Co-treating cells with 100 μM Fe (III) citrate led to a reduction in HIF-1 alpha levels across the three ATP6V0E1 depleted cells. phd2 was knocked down as a control & treatment with 100 μM Fe (III) citrate did not result in reduction of HIF-1 alpha levels (1 × 106 cells per sample harvested & analyzed; N = 2). FACs plot shown is a representative image of two biological repeats performed. (D) Immunoblot analysis for HIF-1 alpha & PHD2 levels in HIF alpha -GFPODD reporter cells with either ATP6V0E1 or PHD2 depleted or treated with 10 nM BafA. The cells were treated with 100 μM Fe (III) citrate for 24 h. beta actin was used as a control. Results validated findings observed by flow cytometry, whereby HIF-1 alpha levels were upregulated following ATP6V0E1 knock-down or inhibition & levels were re-normalized upon Fe (III) citrate treatment. Treatment of Fe (III) citrate in PHD2 depleted cells did not alter HIF-1 alpha levels. All experiments were performed in biological duplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32984302), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - PHD3 depletion stabilizes hypoxic p27 expression by increasing p27 half-life. a Cell cycle arrest at G0 & subsequent release shows an increase of p27 expression in siPHD3 exposed cells. b Quantification for p27 expression under PHD3 depletion at indicated time points after cell cycle release in HeLa & 786-O cells. Asterisk indicates significant difference (p < 0,05; n = 3). c Cell cycle arrest at G0 & inhibition of protein synthesis with cycloheximide indicate increased p27 stability in PHD3 depleted HeLa cells. d Quantification of p27 expression using siPHD3 or control at indicated time points. Four independent experiments (± SEM) are shown (p < 0,05; n = 4). e Analysis of p27 stability in 786–0 cells by cycloheximide chase during reoxygenation after 24 h hypoxia demonstrates markedly increased half-life of p27 upon PHD3 depletion Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pm, Pm-Or, Rb, RM

Species Glossary

Reactivity: Hu, Mu, Rt, Pm, Pm-Or, Rb, RM

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-137AF350
Alexa Fluor 405	NB100-137AF405
Alexa Fluor 488	NB100-137AF488
Alexa Fluor 532	NB100-137AF532
Alexa Fluor 594	NB100-137AF594
Alexa Fluor 647	NB100-137AF647
Alexa Fluor 700	NB100-137AF700
Alexa Fluor 750	NB100-137AF750
Allophycocyanin	NB100-137APC
Biotin	NB100-137B
CoraFluor 1	NB100-137CL1
DyLight 350	NB100-137UV
DyLight 405	NB100-137V
DyLight 488	NB100-137G
DyLight 550	NB100-137R
DyLight 594	NB100-137DL594
DyLight 650	NB100-137C
DyLight 680	NB100-137FR
DyLight 755	NB100-137IR
FITC	NB100-137F
HRP	NB100-137H
Janelia Fluor 525	NB100-137JF525
Janelia Fluor 549	NB100-137JF549
Janelia Fluor 646	NB100-137JF646
Janelia Fluor 669	NB100-137JF669
PE	NB100-137PE
PerCP	NB100-137PCP
mFluor Violet 450 SE	NB100-137MFV450
mFluor Violet 500 SE	NB100-137MFV500
mFluor Violet 610 SE	NB100-137MFV610

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AB-100-NA

TGF-beta Antibody [Unconjugated] - Pan Specific

(95 Publications)

1 mg

AB-100-NA

TGF-beta was detected in immersion fixed paraffin-embedded sections of human prostate tissue using Rabbit Anti-TGF-beta Pan Specific Polyclonal Antibody (Catalog # AB-100-NA) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epitjhelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Porcine TGF-beta 1 (Catalog # 101-B1) inhibits Recombinant Mouse IL-4 (Catalog # 404-ML) induced proliferation in the HT-2 mouse T cell line in a dose-dependent manner (orange line). Inhibition of Recombinant Mouse IL-4 (7.5 ng/mL) activity elicited by Porcine TGF-beta 1 (1 ng/mL) is neutralized (green line) by increasing concentrations of TGF-beta Pan Specific Polyclonal Antibody (Catalog # AB-100-NA). The ND50range is 5-30 µg/mL.

TGF-beta induces IL-9 mRNA expression in Th1 and Th2 cells.Naïve CD4+ T cells were activated with fibroblast-bound anti-CD3/CD28 under classical Th1 and Th2 conditions, with IL-12 and anti-IL-4 or IL-4, anti-IFN-gamma , and anti-IL-12 respectively for 5 days, restimulated at day 5 and split in cultures with and without TGF-beta or anti-TGF-beta for 5 more days of stimulation. (A, C and D) qRT-PCR gene expression analysis of the relative gene expression of IL-9, GATA3, and Tbet at day 10 relative to the Th0 culture. (B) CT value from qRT-PCR gene expression analysis of the gene PU.1. Each donor is represented by a specific symbol and connected with a line. Data are from four independent experiments, each with two donors. *p<0.05, **p<0.01, ***p<0.001. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0021695), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

All-Multi

Species Glossary

Reactivity: All-Multi

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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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

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Protein/Target

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Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

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Promotional Item

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Histone H2AX [p Ser139] Antibody - BSA Free

Rabbit IgG Isotype Control [Unconjugated]

CD31/PECAM-1 Antibody

GFAP Antibody

IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

Rabbit anti-Goat IgG Secondary Antibody [HRP (Horseradish Peroxidase)]

HA Tag Antibody - BSA Free

GAPDH Antibody [Unconjugated]

Stathmin-2/STMN2 Antibody - BSA Free

ZEB1 Antibody

EGLN1/PHD2 Antibody - BSA Free

TGF-beta Antibody [Unconjugated] - Pan Specific