Histone H2AX [p Ser139] Antibody

Novus Biologicals | Catalog # NB100-2280

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Canine

Cited:

Human, Mouse, Canine

Predicted:

Bat (100%), Bovine (100%), Chinese Hamster (100%), Guinea Pig (100%), Monkey (100%), Porcine (100%), Rabbit (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
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Product Specifications

Immunogen

This Histone H2AX [p Ser139] Antibody was developed against a synthetic phospho-peptide, which represented a portion of the C-terminus of human histone H2AX surrounding phosphorylated serine 139 (GeneID 3014).

Reactivity Notes

Based on sequence percent identity: Gorilla (100%), Macaque (100%), Canine reactivity reported in scientific literature (PMID: 26991424).

Modification

p Ser139

Marker

DNA Double-strand break marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

15 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

Novus Biologicals Rabbit Histone H2AX [p Ser139] Antibody (NB100-2280) is a polyclonal antibody validated for use in IHC, WB, ICC/IF and Simple Western. Anti-Histone H2AX Antibody: Cited in 50 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for Histone H2AX [p Ser139] Antibody

gamma-H2AX-[p-Ser139]-Antibody-Immunohistochemistry-NB100-2280-img0022.jpg
Simple Western: Histone H2AX [p Ser139] Antibody [NB100-2280] - 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.
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).
Histone H2AX [p Ser139] Antibody

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/31932580), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/31932580), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/32352929), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/31488102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/32352929), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/30866939), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/32545442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Histone H2AX [p Ser139] Antibody

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-2280] -

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 (https://pubmed.ncbi.nlm.nih.gov/30866939), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Histone H2AX [p Ser139] Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100 - 1:500

Immunohistochemistry-Paraffin

1:100-1:500

Simple Western

1:20

Western Blot

1:100-1:2000
Application Notes

Epitope exposure is recommended. Epitope exposure with citrate buffer will enhance staining. Likely to work with frozen sections. Use in WB reported in scientific literature ( PMID 24415760). Use in IHC-Frozen reported in scientific literature (PMID 26577699).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat lysate, separated by Size, antibody dilution of 1:20, apparent MW was 30 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 5 using NB100-2280 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

TBS and 0.1% BSA

Preservative

0.09% Sodium Azide

Concentration

0.1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: Histone H2AX

Gamma H2AX (gammaH2AX) is the phosphorylated version of histone H2AX and is a marker for double-stranded breaks (DSBs) caused by DNA damage (1-4). H2AX is a variant of histone H2A, one of the histone core molecules forming the nucleosome, and is a vital component in repairing DNA damage (1-4). The H2AX variant represents between 2-25% of total H2A (1-3). Human H2AX protein is 143 amino acids in length and has a theoretical molecular weight of 15 kDa (5). As a result of DSBs, H2AX becomes phosphorylated at serine-139 and gamma H2AX is generated as a response to genomic instability (1-4). Gamma H2AX and its phosphorylation can be detected through western blotting and immunofluorescence, serving as a biomarker for DSB frequency (1-4). Ataxia telangiectasia mutated (ATM) is considered a key protein for H2AX phosphorylation (1-4). Upon the phosphorylation and formation of gamma H2AX, many repair proteins become associated including breast cancer 1 protein (BRCA1), NBS1 (Nijmegen breakage syndrome 1)/hMRE11 (human meiotic recombination 11 protein)/ hRAD50 (N/M/R) repair complex, and 53BP1 (p53 binding protein 1) (1-4). Gamma H2AX detection assays are currently utilized for a variety of purposes, from studying DNA repair to drug development and radiation biodosimetry (4).

References

1. Palla, V. V., Karaolanis, G., Katafigiotis, I., Anastasiou, I., Patapis, P., Dimitroulis, D., & Perrea, D. (2017). gamma-H2AX: Can it be established as a classical cancer prognostic factor?. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine. https://doi.org/10.1177/1010428317695931

2. Kuo, L. J., & Yang, L. X. (2008). Gamma-H2AX - a novel biomarker for DNA double-strand breaks. In vivo (Athens, Greece).

3. Kinner, A., Wu, W., Staudt, C., & Iliakis, G. (2008). Gamma-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic acids research. https://doi.org/10.1093/nar/gkn550

4. Redon, C. E., Weyemi, U., Parekh, P. R., Huang, D., Burrell, A. S., & Bonner, W. M. (2012). gamma-H2AX and other histone post-translational modifications in the clinic. Biochimica et biophysica acta. https://doi.org/10.1016/j.bbagrm.2012.02.021

5. H2AX: Uniprot (P16104)

Alternate Names

H2AFX

Entrez Gene IDs

3014 (Human); 15270 (Mouse)

Gene Symbol

H2AX

UniProt

Additional Histone H2AX Products

Product Documents for Histone H2AX [p Ser139] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Histone H2AX [p Ser139] Antibody

Licensed to Novus Biologicals LLC under U.S. Patent Nos. 6,362,317 and 6,884,873.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Citations for Histone H2AX [p Ser139] Antibody

Customer Reviews for Histone H2AX [p Ser139] Antibody (1)

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  • Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Ramos lymphoma
    Species: Human
    Verified Customer | Posted 10/06/2015
    Lymphoma treated with DNA damaging agent
    Histone H2AX [p Ser139] Antibody NB100-2280

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FAQs for Histone H2AX [p Ser139] Antibody

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    • Q: Could you direct me where to find secondary antibodies for NB100-2280 that would be appropriate for certain applications?

      A: By clicking on the "Support & Research" tab on NB100-2280's product page under the "Product Details" display, you will find a link to view all anti-Rabbit secondary antibodies. This will lead you to a new page that will allow you to filter products by application.
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