Anti-Histone H2AX Antibodies: 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>gamma-H2AX-[p-Ser139]-Antibody-Immunohistochemistry-NB100-384-img0024.jpg

gamma-H2AX-[p-Ser139]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-384-img0022.jpg

<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.

<b>Biological Strategies Validation. </b>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.

<b>Biological Strategies Validation. </b>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
Alexa Fluor Plus 405	NB100-384AFP405
Alexa Fluor Plus 488	NB100-384AFP488
Alexa Fluor Plus 555	NB100-384AFP555
Alexa Fluor Plus 594	NB100-384AFP594
Alexa Fluor Plus 647	NB100-384AFP647
Alexa Fluor Plus 680	NB100-384AFP680
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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NB100-2280

Histone H2AX [p Ser139] Antibody

(1 Review)

(54 Publications)

0.1 ml

NB100-2280

gamma-H2AX-[p-Ser139]-Antibody-Immunohistochemistry-NB100-2280-img0021.jpg

Immunohistochemistry-Paraffin: Histone H2AX [p Ser139] Antibody [NB100-2280] - Detection of human Histone H2AX [p Ser139] antibody by immunohistochemistry. Sample: FFPE section of human seminoma. Antibodies: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at a dilution of 1:500. Detection: DAB.

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-2280] - Detection of Mouse Histone H2AX [p Ser139] by Immunohistochemistry. Sample: FFPE section of mouse colon carcinoma CT26. Antibodies: Affinity purified rabbit Histone H2AX [p Ser139] antibody. Detection: DAB.

Simple Western: Histone H2AX [p Ser139] Antibody [NB100-2280] - Electropherogram image(s) of corresponding Simple Western lane view. Histone H2AX [p Ser139] antibody was used at 1:20 dilution on Jurkat lysate(s).

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] - Treatment with anti-thyroid drug protected RPE & photoreceptors from oxidative damage induced by NaIO3.RPE & retinal oxidative damage were evaluated by immunofluorescence labeling of p-gamma H2AX & 8-OHdG on the RPE whole mounts & retinal sections at 3 days post-NaIO3 injection. a Shown are representative images of p-gamma H2AX immunofluorescence labeling on the RPE whole mounts. b Shown are representative images of p-gamma H2AX & 8-OHdG immunofluorescence labeling on the retinal sections, & corresponding quantitative analysis for p-gamma H2AX labeling. ONL, outer nuclear layer; INL, inner nuclear layer. Data represented the mean ± SEM for 5 mice per group (*p < 0.05). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31932580), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] - RES rescued DXR-induced apoptosis through DNA-damage-P63-Caspase3 pathway in mouse oocytes. (A) Representative immunofluorescence images showing the expression of gamma -H2AX in mouse oocytes. Green, gamma -H2AX, Blue, DNA, Bar = 20 μm. (B) The relative immunofluorescence intensity of gamma -H2AX was measured in control, DXR-treated & RES-supplemented oocytes. Experiments were repeated at least 3 times with more than 30 oocytes examined for each group. Data were presented as means ± S.E.M of three independent experiments. **means P < 0.01, *** means P < 0.001. (C) Protein levels of gamma -H2AX, P63 & Active-Caspase3 were examined by Western blotting in control, DXR-treated & RES-supplemented oocytes. GAPDH was used as a loading control. The clean backgrounds for the active-Caspase-3, gamma -H2AX & GAPDH is due to the exposure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32352929), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-2280] - Hepatic activation of FOXO3 induces oxidative damage & Akt activation. a Expression of Bcl2l11 (Bim), Sesn2 & Sesn3 by qPCR in livers from 9-week-old control & FOXO3CAHep transgenic mice (n = 4). b Representative IHC staining for 8-OHdG (bar = 100 μm) & quantification of 8-OHdG-positive cells as a percentage of total hepatocyte cells in livers of patients in area of small cells (SC) & large cells (LC). c Representative IHC staining for phospho-gamma H2AX (bar = 100 μm) & quantification of phospho-gamma H2AX-positive cells as a percentage of total hepatocyte cells in livers of patients in area of small cells & large cells. d Expression of Msh2 by qPCR (upper panel), Western blot analysis for MSH2 (middle panel) & densitometric quantification (lower panel) in livers from 9-week-old control & FOXO3CAHep transgenic mice (n = 4). e Western blot analysis in livers from 9-week-old control & FOXO3CAHep mice for serine-473 phospho-Akt, with Akt & GAPDH as loading control (left panel), as well as densitometric quantification (right panel). f Representative IHC staining for phospho-Akt in livers from 9-week-old control & FOXO3CAHep mice (bar = 100 μm). g Western blot analysis in livers from 9-week-old control & FOXO3CAHep mice for Rictor & GAPDH as loading control (left panel), as well as densitometric quantification (right panel) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31488102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-2280] - RES rescued DXR-induced apoptosis through DNA-damage-P63-Caspase3 pathway in mouse oocytes. (A) Representative immunofluorescence images showing the expression of gamma -H2AX in mouse oocytes. Green, gamma -H2AX, Blue, DNA, Bar = 20 μm. (B) The relative immunofluorescence intensity of gamma -H2AX was measured in control, DXR-treated & RES-supplemented oocytes. Experiments were repeated at least 3 times with more than 30 oocytes examined for each group. Data were presented as means ± S.E.M of three independent experiments. **means P < 0.01, *** means P < 0.001. (C) Protein levels of gamma -H2AX, P63 & Active-Caspase3 were examined by Western blotting in control, DXR-treated & RES-supplemented oocytes. GAPDH was used as a loading control. The clean backgrounds for the active-Caspase-3, gamma -H2AX & GAPDH is due to the exposure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32352929), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] - Oocyte-specific deletion of Gsk-3 beta disrupted early folliculogenesis in mice. aGsk-3 beta efficiently & specifically deleted in the oocytes in cKO mouse ovary. Immunofluorescence staining for GSK-3 beta (green) & DDX4 (red) on 1 dpp ovary. The nucleus stained by Hoechst (blue). b Control & cKO ovaries at the indicated developmental stages. Oocytes stained w/ DDX4 (green). The nucleus stained by Hoechst (blue). c Statistical analysis showed that the total number of primordial follicle decreased significantly in 7 dpp cKO ovaries (Additional file 8: Individual data values). d Apoptotic cells increased in 1 dpp cKO ovaries compared w/ the control ovaries. TUNEL signals (green) marked apoptotic cells. The nucleus stained by Hoechst (blue). e DSBs persisted in the oocytes of 1 dpp cKO ovaries. The sections stained w/ gamma -H2AX (green) & DDX4 (red). The nucleus stained by Hoechst (blue). f Ectopic RAD51 expression in the oocytes of 1 dpp cKO ovaries. The sections stained w/ RAD51 (green) & DDX4 (red). The nucleus stained by Hoechst (blue). g, h beta -catenin accumulated in the cytoplasm & translocated into nucleus of the oocytes in cKO mice. g The sections stained w/ beta -catenin (green) & DDX4 (red). The nucleus stained by Hoechst (blue). Arrowheads indicated oocytes showing nuclear beta -catenin accumulation. h The statistic analysis demonstrated that the percentage of oocytes showing beta -catenin accumulation per section increased significantly in cKO mice (Additional file 8: Individual data values). The data are presented as mean ± s.d. The asterisk (*) denotes a statistically significant difference between the control & treatment groups. *P < 0.05, **P < 0.01, & ***P < 0.001 (t test). Scale bars, 200 μm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30866939), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-2280] - IDO1 inhibitor enhances the efficacy of radiotherapy in vivo. 1 × 105 of SiHa tumorsphere cells were subcutaneously injected to nude mice for tumor growth. After the tumor volume reached 50 mm3, the mice were divided into four groups of non-treated (Ctrl), INCB-024360 treated (INCB), radiotherapy (Rx), or INCB-024360 plus radiotherapy (INCB + Rx). For the INCB or INCB + Rx group, mice were injected once with 50 mg/kg INCB-024360 intraperitoneally before radiotherapy. For the Rx or INCB + Rx group, mice received 2 Gy radiation per day for total 10 Gy. Mice were sacrificed at day 30 after the last radiation treatment & the xenografted tumors were taken out for picturing (A) & weighting (B). The expression of Ki-67 or p-gamma H2AXser139 was determined by paraffin section followed by immunohistochemical staining (C). The inserted bars indicated 50 μM. The quantification results were performed by TissueFAX software (D). * p < 0.05; ** p < 0.01. The experiments were repeated two times & data from one experiment were presented. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32545442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] - Premature upregulation of TAp63 in GSK-3 beta -inhibited ovary. a Expression pattern of gamma -H2AX in fetal & neonatal mouse ovary in vivo. Mouse ovaries from 13.5 dpc, 15.5 dpc, 17.5 dpc, & 1 dpp immunostained for gamma -H2AX (green) & DDX4 (red). Nucleus stained by Hoechst (blue). gamma -H2AX displayed intensive expression in germ cell nucleus from 15.5 to 17.5 dpc. b Expression pattern of TAp63 in fetal & neonatal mouse ovary in vivo. Mouse ovaries from 13.5 dpc, 15.5 dpc, 17.5 dpc, 18.5 dpc, & 1 dpp immunostained for TAp63 (green) & DDX4 (red). Nucleus stained by Hoechst (blue). TAp63 protein located w/in somatic cells in fetal ovary & began to express in germ cell nucleus from 18.5 dpc afterward. c–e TAp63 expression upregulated in fetal ovary & displayed premature localization w/in oocyte nucleus following GSK-3 beta inhibition. Before examination, ovaries of 14.5 dpc cultured in vitro w/ DMSO or BIO for 3 days. c qRT-PCR analysis of TAp63 mRNA level following GSK-3 beta inhibition (normalized to beta -actin) (Additional file 8: Individual data values). d WB analysis of TAp63 protein level following GSK-3 beta inhibition. GAPDH used as internal control. e Immunofluorescence staining for TAp63 (green) & DDX4 (red). The nucleus stained by Hoechst (blue). TAp63 showed premature expression w/in oocyte nucleus following GSK-3 beta inhibition (arrowhead). f qRT-PCR results showed that relative mRNA expression level of p21, Bad, & Noxa increased significantly in GSK-3 beta -inhibiting ovaries (Additional file 8: Individual data values). data are presented as mean ± s.d. The asterisk (*) denotes a statistically significant difference between the control & treatment groups. *P < 0.05, **P < 0.01, & ***P < 0.001 (t test). Scale bars, 200 μm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30866939), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Ca, Po, Bt, Bv, ChHa, Gp, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Ca, Po, Bt, Bv, ChHa, Gp, Pm, Rb

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NB100-74435

Histone H2AX [p Ser139] Antibody (3F2)

(1 Review)

(22 Publications)

100 μg

NB100-74435

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - Staining using NB100-74435, treatment with paraquat and iron induces MnSOD and Phosphorylation of H2AX in RAW 264.7 macrophages. Cells were treated for 20 hours with paraquat (500 uM) and iron (200 ug/ml) and stained with anti-Phospho-H2AX antibody.

<b>Biological Strategies Validation. </b>Western Blot: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - Western blot analysis of Jurkat cells (Lane 1) and Jurkat cells treated with staurosporine (Lane 2) blotted with Histone H2AX [p Ser139] antibody (3F2). Bands indicate an observed molecular weight of ~16 kDa.

Immunocytochemistry/Immunofluorescence: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - STED image of MDA-MB-231 cells after 2 Gy irradiation. Blue channel: DAPI, window level [0 255]. Green channel: secondary antibody Alexa Fluor 488, window level [140 255]. Scale bar: 20 um. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - Immunofluorescence analysis using Histone H2AX [p Ser139] antibody (3F2).

Immunocytochemistry/Immunofluorescence: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - IF image of phosphorylated H2AX in gamma irradiated Hela cells.

Simple Western: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - Electropherogram image(s) of corresponding Simple Western lane view. Histone H2AX [p Ser139] antibody (3F2) was used at 10 ug/ml dilution on Jurkat lysate(s).

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - Evidence of DNA damage & oxidative stress in islets from HFD treated mice. (a) Representative immunofluorescent staining of gamma H2AX, insulin, & DAPI for islets from each treatment group. (b) Representative immunofluorescent staining of 4-hydroxynonenal (4-HNE), insulin, & DAPI for islets from each treatment group. (c) Representative immunofluorescent staining of nitrotyrosine, insulin, & DAPI for islets from each treatment group. (d–g) qRT-PCR analysis of total islet RNA for antioxidant genes, including Gpx1, SOD1, Nrf2, & Ppargc1 alpha . (n = 3) *p < 0.05, **p < 0.01. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-03869-5), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Bv

Species Glossary

Reactivity: Hu, Mu, Bv

Applications:

, , , , ,

Applications: , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

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NB100-638

Histone H2AX Antibody - BSA Free

(29 Publications)

100 ul

NB100-638

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Immunocytochemistry/Immunofluorescence: Histone H2AX Antibody [NB100-638] - Asynchronous wild-type (+/+) and H2AX knockout (-/-) mouse embryonic fibroblasts. Antibody used at 2 ug/ml.

<b>Genetic Strategies Validation. </b>Western Blot: Histone H2AX Antibody [NB100-638] - Human 293T and wild-type (+/+) or H2AX knockout (-/-) mouse embryonic fibroblasts. Antibody used at the indicated concentrations.

Histone-H2AX-Antibody-Western-Blot-NB100-638-img0005.jpg

<b>Biological Strategies Validation. </b>Histone-H2AX-Antibody-Western-Blot-NB100-638-img0007.jpg

Western Blot: Histone H2AX Antibody [NB100-638] - Knockdown of CAPER induces DSB proteins ATM & H2AX & leads to apoptosis in MDA-MB-231 cells(A) H2AX phosphorylation on ser139 is significantly increased (upper right quadrant population) in MDA-MB-231 cells after CAPER knockdown as represented by Muse Cell Analyzer plots (3-fold, p < 0.001, n = 3). (B) The increase in gamma H2AX was also validated through western blot analysis (2.5-fold, p < 0.01, n = 3). ATM phosphorylation on serine1981 is significantly upregulated after knockdown of CAPER expression (5-fold, p < 0.05, n = 3). (C) CAPER knockdown resulted in an increased level of caspase-3/7 activation through decreasing live cells (1.2-fold, p < 0.001, n = 3), while increasing apoptotic (7.5-fold, p < 0.001, n = 3), apoptotic/dead (6.5-fold, p < 0.001, n = 3), & dead (3-fold, p < 0.05, n = 3) cell populations. Interestingly, MDA-MB-231 cells expressing CAPER knockdown displayed no significant changes in any of the phases of the cell cycle (p = NS, n = 8, for G1, S & G2/M phases) compared to CTL shRNA (Figure 7C right panels). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30100993), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-638AF350
Alexa Fluor 405	NB100-638AF405
Alexa Fluor 488	NB100-638AF488
Alexa Fluor 532	NB100-638AF532
Alexa Fluor 594	NB100-638AF594
Alexa Fluor 647	NB100-638AF647
Alexa Fluor 700	NB100-638AF700
Alexa Fluor 750	NB100-638AF750
Alexa Fluor Plus 405	NB100-638AFP405
Alexa Fluor Plus 488	NB100-638AFP488
Alexa Fluor Plus 555	NB100-638AFP555
Alexa Fluor Plus 594	NB100-638AFP594
Alexa Fluor Plus 647	NB100-638AFP647
Alexa Fluor Plus 680	NB100-638AFP680
Allophycocyanin	NB100-638APC
Biotin	NB100-638B
CoraFluor 1	NB100-638CL1
DyLight 350	NB100-638UV
DyLight 405	NB100-638V
DyLight 488	NB100-638G
DyLight 550	NB100-638R
DyLight 594	NB100-638DL594
DyLight 650	NB100-638C
DyLight 680	NB100-638FR
DyLight 755	NB100-638IR
FITC	NB100-638F
HRP	NB100-638H
Janelia Fluor 525	NB100-638JF525
Janelia Fluor 549	NB100-638JF549
Janelia Fluor 646	NB100-638JF646
Janelia Fluor 669	NB100-638JF669
PE	NB100-638PE
PerCP	NB100-638PCP
mFluor Violet 450 SE	NB100-638MFV450
mFluor Violet 500 SE	NB100-638MFV500
mFluor Violet 610 SE	NB100-638MFV610

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AF2288

Histone H2AX [p Ser139] Antibody [Unconjugated]

(28 Publications)

50 μg

AF2288

CTS013). Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox">

Histone H2AX was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # <a class=

CTS013). Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." alt="Histone H2AX was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL overnight at 4 °C. 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 the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

HAF008). A specific band was detected for Phospho-Histone H2AX (S139) at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 µM camptothecin (CPT) for for the indicated times. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # <a class=

HAF008). A specific band was detected for Phospho-Histone H2AX (S139) at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

<b>Biological Strategies Validation. </b>Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 µM Camptothecin (CPT) for 24 hours, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Histone H2AX (S139) at approximately 25 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

<b>Biological Strategies Validation. </b>Histone H2AX phosphorylated at S139 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with ultraviolet radiation using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # <a class=

NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

Meflufen induces DNA damage and apoptosis in cancer cells. (C) D492, D492HER2, and MDA‐MB231 cells were incubated with 1 µmol L‐1 of melflufen for 30 min and harvested on indicated timepoints and subjected for immunoblotting for H2AX/ yH2Ax. One representative blot of 3 is shown. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32717133), licensed under a CC-BY license. Not internally tested by R&D Systems.

Meflufen induces DNA damage and apoptosis in cancer cells. (A) D492, D492HER2, and MDA‐MB231 cells were seeded on a six well plate and incubated with either none, 1 µmol L‐1 or 5 µmol L‐1 melflufen for 30 min and harvested 3 h post‐treatment. Equally loaded protein lysates subjected for immunoblotting for H2AX/ yH2Ax One representative blot of three is shown. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32717133), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2288-SP

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NB100-383

Histone H2AX Antibody - BSA Free

(14 Publications)

100 ul

NB100-383

Western Blot: Histone H2AX Antibody [NB100-383] - Detection of mouse H2AX by western blot. Samples: Whole cell lysate (50 ug) from F9 and L1210 cells prepared using RIPA lysis buffer. Antibody: Affinity purified rabbit anti-H2AX antibody A300-083A (lot A300-083A-5) used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

Simple Western: Histone H2AX Antibody [NB100-383] - Simple Western lane view shows a specific band for H2AX in 0.2 mg/ml of HEK lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa separation system.

Simple Western: Histone H2AX Antibody [NB100-383] - Electropherogram image(s) of corresponding Simple Western lane view. H2AX antibody was used at 10 ug/ml dilution on HEK lysate(s).

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-383AF350
Alexa Fluor 405	NB100-383AF405
Alexa Fluor 488	NB100-383AF488
Alexa Fluor 532	NB100-383AF532
Alexa Fluor 594	NB100-383AF594
Alexa Fluor 647	NB100-383AF647
Alexa Fluor 700	NB100-383AF700
Alexa Fluor 750	NB100-383AF750
Alexa Fluor Plus 405	NB100-383AFP405
Alexa Fluor Plus 488	NB100-383AFP488
Alexa Fluor Plus 555	NB100-383AFP555
Alexa Fluor Plus 594	NB100-383AFP594
Alexa Fluor Plus 647	NB100-383AFP647
Alexa Fluor Plus 680	NB100-383AFP680
Allophycocyanin	NB100-383APC
Biotin	NB100-383B
CoraFluor 1	NB100-383CL1
DyLight 350	NB100-383UV
DyLight 405	NB100-383V
DyLight 488	NB100-383G
DyLight 550	NB100-383R
DyLight 594	NB100-383DL594
DyLight 650	NB100-383C
DyLight 680	NB100-383FR
DyLight 755	NB100-383IR
FITC	NB100-383F
HRP	NB100-383H
Janelia Fluor 525	NB100-383JF525
Janelia Fluor 549	NB100-383JF549
Janelia Fluor 646	NB100-383JF646
Janelia Fluor 669	NB100-383JF669
PE	NB100-383PE
PerCP	NB100-383PCP
mFluor Violet 450 SE	NB100-383MFV450
mFluor Violet 500 SE	NB100-383MFV500
mFluor Violet 610 SE	NB100-383MFV610

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NBP2-27428

Histone H2AX Antibody

(1 Publication)

0.1 mg

NBP2-27428

Immunohistochemistry-Paraffin: H2AX Antibody [NBP2-27428] - Formalin-fixed, paraffin-embedded human spleen stained with Histone H2A.x antibody at 10 ug/ml.

Immunocytochemistry/Immunofluorescence: H2AX Antibody [NBP2-27428] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-H2AX at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Alt Catalog#

IMG-6593A

Alt Catalog# IMG-6593A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

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MAB3406

Histone H2AX Antibody (322105)

(11 Publications)

50 μg

MAB3406

HAF007). A specific band was detected for Histone H2AX at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

Western blot shows lysates of HEK293 human embryonic kidney cell line, K562 human chronic myelogenous leukemia cell line, Balb/3T3 mouse embryonic fibroblast cell line, and Nb2-11 rat lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat Histone H2AX Monoclonal Antibody (Catalog # MAB3406) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # <a class=

HAF007). A specific band was detected for Histone H2AX at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HEK293 human embryonic kidney cell line, K562 human chronic myelogenous leukemia cell line, Balb/3T3 mouse embryonic fibroblast cell line, and Nb2-11 rat lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat Histone H2AX Monoclonal Antibody (Catalog # MAB3406) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Histone H2AX at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

Histone H2AX was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat Histone H2AX Monoclonal Antibody (Catalog # MAB3406) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # <a class=

NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

The NAMPT-dependent NAD+ salvage pathway is necessary for the recovery of NAD+ & induction of apoptosis under weak H2O2 stimulation. A The amounts of endogenous gamma H2AX & NAMPT proteins under 0.7 & 1.1 mM H2O2 stimulation determined by immunoblotting (n = 3). Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35410407), 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:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB3406-SP

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NBP1-19255

Histone H2AX [p Ser139] Antibody (9F3) - BSA Free

(4 Publications)

0.05 mg

NBP1-19255

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Ch, Gp, Ha, Pm, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Ch, Gp, Ha, Pm, Rb, Sh

Applications:

, ,

Applications: , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-19255-0.2mg

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NBP3-14678

Histone H2AX [p Ser139] Antibody (BLR053F)

100 ul

NBP3-14678

Immunohistochemistry-Paraffin: Histone H2AX Antibody (BLR053F) [NBP3-14678] - Section of human breast carcinoma. Antibody: Rabbit Histone H2AX antibody (BLR053F) used at 1:100. Secondary: DyLight(R) 594-conjugated goat anti-rabbit IgG. Counterstain: DAPI.

Flow Cytometry: Histone H2AX Antibody (BLR053F) [NBP3-14678] - Rabbit Histone H2AX antibody (BLR053F) or isotype control (unshaded). Secondary: DyLight(R) 488-conjugated goat anti-rabbit IgG.

Immunohistochemistry-Paraffin: Histone H2AX Antibody (BLR053F) [NBP3-14678] - Section of human basal cell carcinoma. Antibody: Rabbit Histone H2AX antibody (BLR053F) used at 1:250. Secondary: HRP-conjugated goat anti-rabbit IgG. Substrate:DAB.

Immunohistochemistry-Paraffin: Histone H2AX Antibody (BLR053F) [NBP3-14678] - Section of mouse CT26 tumor. Antibody: Rabbit Histone H2AX antibody (BLR053F) used at 1:250. Secondary: HRP-conjugated goat anti-rabbit IgG. Substrate:DAB.

<b>Biological Strategies Validation. </b>Immunoprecipitation: Histone H2AX Antibody (BLR053F) [NBP3-14678] - Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from Jurkat cells treated with 100 uM etoposide (+) or mock treated (-). Antibodies: Rabbit anti-gamma-H2AX recombinant monoclonal antibody used for IP at 20 ul per reaction. gamma-H2AX was also immunoprecipitated by rabbit anti-gamma-H2AX antibody. For blotting immunoprecipitated gamma-H2AX, was used at 1:1000. Chemiluminescence with an exposure time of 3 seconds. Lower panel: Rabbit anti-H2AX

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NBP3-15378

Histone H2AX Antibody (4Z3S8)

100 ul

NBP3-15378

Immunohistochemistry-Paraffin: Histone H2AX Antibody (4Z3S8) [NBP3-15378] - Immunohistochemistry of paraffin-embedded Human liver cancer using Histone H2AX antibody (NBP3-15378) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

Immunohistochemistry-Paraffin: Histone H2AX Antibody (4Z3S8) [NBP3-15378] - Immunohistochemistry of paraffin-embedded Rat brain using Histone H2AX antibody (NBP3-15378) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

Chromatin Immunoprecipitation: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Chromatin immunoprecipitation analysis of extracts of HeLa cells, using Histone H2AX Rabbit mAb antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Human cervix cancer tissue using Histone H2AX Rabbit mAb at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Mouse intestin tissue using Histone H2AX Rabbit mAb at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Human colon tissue using Histone H2AX Rabbit mAb at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using Histone H2AX Rabbit mAb at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using Histone H2AX Rabbit mAb at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Rat brain tissue using Histone H2AX Rabbit mAb at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using Histone H2AX Rabbit mAb at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Western Blot: Histone H2AX Antibody (4Z3S8) [Histone H2AX] - Western blot analysis of various lysates using Histone H2AX Rabbit mAb at 1:1000 dilution. <br>Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution. <br>Lysates/proteins: 25 ug per lane.<br>Blocking buffer: 3% nonfat dry milk in TBST.<br>Detection: ECL Basic Kit .<br>Exposure time: 10s.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, ,

Applications: , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Conjugate:

Unconjugated

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NBP3-04113

Histone H2AX Antibody - BSA Free

100 ul

NBP3-04113

Immunocytochemistry/ Immunofluorescence: Histone H2AX Antibody - BSA Free [Histone H2AX] - Confocal immunofluorescence analysis of HeLa cells using Histone H2AX Rabbit pAb at dilution of 1:200. Blue: DAPI for nuclear staining.

Immunohistochemistry: Histone H2AX Antibody - BSA Free [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Rat heart using Histone H2AX Rabbit pAb at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody - BSA Free [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Mouse brain using Histone H2AX Rabbit pAb at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

Immunohistochemistry: Histone H2AX Antibody - BSA Free [Histone H2AX] - Immunohistochemistry analysis of paraffin-embedded Rat brain using Histone H2AX Rabbit pAb at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

Western Blot: Histone H2AX Antibody - BSA Free [Histone H2AX] - Western blot analysis of various lysates using Histone H2AX Rabbit pAb at 1:1000 dilution.<br>Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.<br>Lysates/proteins: 25ug per lane.<br>Blocking buffer: 3% nonfat dry milk in TBST.<br>Detection: ECL Basic Kit .<br>Exposure time: 90s.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP3-04113-20ul

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

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

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Clone

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Species Glossary

Histone H2AX Antibodies

Histone H2AX [p Ser139] Antibody - BSA Free

Histone H2AX [p Ser139] Antibody

Histone H2AX [p Ser139] Antibody (3F2)

Histone H2AX Antibody - BSA Free

Histone H2AX [p Ser139] Antibody [Unconjugated]

Histone H2AX Antibody - BSA Free

Histone H2AX Antibody

Histone H2AX Antibody (322105)

Histone H2AX [p Ser139] Antibody (9F3) - BSA Free

Histone H2AX [p Ser139] Antibody (BLR053F)

Histone H2AX Antibody (4Z3S8)

Histone H2AX Antibody - BSA Free