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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Human, Mouse, Rat, Canine

Applications

Chromatin Immunoprecipitation (ChIP), Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

1.0 mg/ml

Product Summary for Histone H2AX [p Ser139] Antibody

Immunogen

This Histone H2AX [p Ser139] Antibody was developed against to a region surrounding phosphorylated serine 139 of human histone H2AX [Swiss-Prot entry P16104] (GeneID 3014).

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 27102221), Canine reactivity reported in scientific literature (PMID: 23365434).

Modification

p Ser139

Localization

Nucleous, chromosome.

Specificity

The epitope maps to a region surrounding phosphorylated serine 139 of human histone H2AX.

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.

Scientific Data Images for Histone H2AX [p Ser139] Antibody

Knockout Validation of Histone H2AX Antibody in Human and Mouse Cells in Western Blot

Knockout Validation of Histone H2AX Antibody in Human and Mouse Cells in Western Blot

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.
Immunocytochemistry/Immunofluorescence Staining of Histone H2AX in Treated and Untreated HeLa Cells

Immunocytochemistry/Immunofluorescence Staining of Histone H2AX in Treated and Untreated HeLa Cells

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 Analysis of Histone H2AX in Jurkat Cell Lysate

Simple Western Analysis of Histone H2AX in Jurkat Cell Lysate

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.

Applications for Histone H2AX [p Ser139] Antibody

Application
Recommended Usage

Flow Cytometry

5 ug per 1 million cells

Immunocytochemistry/ Immunofluorescence

1:500 to 1:5000

Immunohistochemistry

1:2000 - 1:10000

Immunohistochemistry-Frozen

1:1000 - 1:5000

Immunohistochemistry-Paraffin

1:2000 - 1:10000

Simple Western

5 ug/ml

Western Blot

1:10000-1:25000
Application Notes
For IHC, epitope retrieval with citrate buffer pH6.0 is recommended for FFPE tissue sections. Formaldehyde fixation is recommended. Permeabilization with Triton-X 100 is recommended for formaldehydefixed cells. Immunoprecipitation is not recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. Use in chromatin immunoprecipitation reported in scientific literature (PMID: 30049290).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using NB100-384 in the following applications:

Published Applications

Read 158 publications using NB100-384 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Citrate/Phosphate (pH 7.0 - 8.0)

Preservative

0.09% Sodium Azide

Concentration

1.0 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

Product Documents for Histone H2AX [p Ser139] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot 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.

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FAQs for Histone H2AX [p Ser139] Antibody

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  • Q: Can we apply antibody gamma H2AX [p Ser139] Antibody (NB100-384) (0.1 ml) for drosophila?

    A: NB100-384 has been validated for use in human and mouse, it has not been tested nor validated for use in Drosophila. If you would be interested in testing this novel species, please take a look at our Innovators Reward Program.

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