Histone H2AX [p Ser139] Antibody (3F2)

Novus Biologicals | Catalog # NB100-74435

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Bovine

Cited:

Human, Mouse, Rabbit

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Immunomicroscopy, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 3F2
View all Histone H2AX Primary Antibodies »

Product Specifications

Immunogen

This Histone H2AX [p Ser139] Antibody (3F2) was developed against a synthetic peptide sequence surrounding phosphorylated Ser139.

Reactivity Notes

Bovine reactivity reported in scientific literature (PMID: 17604361). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Modification

p Ser139

Marker

DNA Double-strand break marker

Specificity

In Western blot this antibody detects ~17 kDa protein representing phosphorylated H2AX in gamma irradiated HeLa cell lysate. In immunofluorescence procedures, recognizes phosphorylated H2AX in gamma irradiated HeLa cells. ELISA of phosphorylated H2AX can also be performed. Used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

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.

Scientific Data Images for Histone H2AX [p Ser139] Antibody (3F2)

Simple Western: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - 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.
Immunocytochemistry/Immunofluorescence: Histone H2AX [p Ser139] Antibody (3F2) [NB100-74435] - Staining of Phospho-H2AX in A549 cells. Cells were treated with 50 uM EPE for 1 hour (left panel) or with vehicle (right control: 0.1% DMSO in media).
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.
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).
Histone H2AX [p Ser139] Antibody (3F2)

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

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 (https://www.nature.com/articles/s41598-017-03869-5), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Histone H2AX [p Ser139] Antibody (3F2)

Application
Recommended Usage

ELISA

1:100 - 1:2000

Flow Cytometry

1 ug/million cells

Immunocytochemistry/ Immunofluorescence

2 - 4 ug/ml

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Paraffin

1:10 - 1:500

Simple Western

10 ug/ml

Western Blot

1 ug/ml
Application Notes
In WB: Detects an approx. 17 kDa protein representing phosphorylated H2AX in gamma irradiate Hela cell lysate.

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 10 ug/mL, apparent MW was 28 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 2 reviews rated 5 using NB100-74435 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS with 1 mg/ml BSA

Preservative

0.05% Sodium Azide

Concentration

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 -20C. Avoid freeze-thaw cycles.

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

P16104

Additional Histone H2AX Products

Product Documents for Histone H2AX [p Ser139] Antibody (3F2)

Certificate of Analysis

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Product Specific Notices for Histone H2AX [p Ser139] Antibody (3F2)

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

Epigenetics

Citations for Histone H2AX [p Ser139] Antibody (3F2)

Customer Reviews for Histone H2AX [p Ser139] Antibody (3F2) (2)

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Customer Images

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  • Excellent antibody for WB
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Colon tissue
    Species: Mouse
    Verified Customer | Posted 10/02/2024
    Dual pink protein marker
    10 mg loading 1:1000 dilution HRP detection
    Histone H2AX [p Ser139] Antibody (3F2) NB100-74435
  • gamma H2AX [p Ser139] Antibody (3F2)
    Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: MDA MB 231 cells
    Species: Human
    Verified Customer | Posted 05/17/2022
    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 μm.
    Histone H2AX [p Ser139] Antibody (3F2) NB100-74435

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