Histone H2AX [p Ser139] Antibody (2F3)

Product Discontinued

Histone H2AX [p Ser139] Antibody (2F3) Summary

• Immunogen: Modified peptide (KATQASQEY)
• Modification: p Ser139
• Marker: DNA Double-strand break marker
• Specificity: H2A.X (2F3) [Ser139]
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: H2AX
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:10-1:1000
  • Immunocytochemistry/ Immunofluorescence 1-4 ug/ml
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Western Blot 1 ug/ml
• Application Notes: Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 5 mug antibody per 5 ml antibody dilution buffer for each mini-gel. For immunofluorescence microscopy: Use a dilution range of 1~4 microgram/ml. It is recommended that the reagent be titrated for optimal performance for each application. FC reported in literature (PMID:23449797).
• Theoretical MW: 15.145 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.
• Control:

  Histone H2AX Overexpression Lysate

Reactivity Notes

Reacts with Ser139-phosphorylated H2A.X. 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.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS (pH 7.2)
• Preservative: 0.09% Sodium Azide
• Concentration: 0.5 mg/ml
• Purity: Protein A or G purified

Notes

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

Alternate Names for Histone H2AX [p Ser139] Antibody (2F3)

• H2A.X
• H2A/X
• H2AFX
• Histone H2AX

Background

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)

Limitations

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.

Publications for gamma H2AX Antibody (NB100-78356)(36)

Publications using NB100-78356

• Ciminera AK, Shuck SC, Termini J Elevated glucose increases genomic instability by inhibiting nucleotide excision repair Life science alliance 2021-10-01 [PMID: 34426491] (WB, ICC/IF, Human)
• Zastko L, Petrovicova P, Rackova A et al. DNA damage response and apoptosis induced by hyperthermia in human umbilical cord blood lymphocytes Toxicology in vitro : an international journal published in association with BIBRA 2021-02-27 [PMID: 33652125] (ICC/IF, Human)
• Alvarez-Palomo AB, Requena-Osete J, Delgado-Morales R et al. A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair generating improved genetically stable human induced pluripotent stem cells Stem cells (Dayton, Ohio) 2021-02-23 [PMID: 33621399] (ICC/IF, Mouse)
• Konkova M, Abramova M, Kalianov A et al. Mesenchymal Stem Cells Early Response to Low-Dose Ionizing Radiation Frontiers in Cell and Developmental Biology 2020-12-14 [PMID: 33381502]
• Tsai LJ, Lopezcolorado FW, Bhargava R et al. RNF8 has both KU-dependent and independent roles in chromosomal break repair Nucleic Acids Res. 2020-05-19 [PMID: 32427332]
• Lagund in D, Hu W, Law H et al. Delineating the role of FANCA in glucose-stimulated insulin secretion in beta cells through its protein interactome PLoS ONE 2019-08-28 [PMID: 31461451]
• Hu WF, Krieger KL, Lagundzin D et al. CTDP1 regulates breast cancer survival and DNA repair through BRCT-specific interactions with FANCI Cell Death Discov 2019-06-19 [PMID: 31240132] (ICC/IF, Human)
• Smith R, Sellou H, Chapuis C et al. CHD3 and CHD4 recruitment and chromatin remodeling activity at DNA breaks is promoted by early poly(ADP-ribose)-dependent chromatin relaxation Nucleic Acids Res. 2018-05-04 [PMID: 29733391] (ICC/IF, Human)
• Onyango DO, Lee G, Stark JM. PRPF8 is important for BRCA1-mediated homologous recombination Oncotarget. 2017-10-06 [PMID: 29212152] (ICC/IF, Human)
• Tran TQ, Ishak Gabra MB, Lowman XH et al. Glutamine deficiency induces DNA alkylation damage and sensitizes cancer cells to alkylating agents through inhibition of ALKBH enzymes PLoS Biol. 2017-11-01 [PMID: 29107960] (ICC/IF, Mouse)
• Sergeeva VA, Ershova ES, Veiko NN et al. Low-Dose Ionizing Radiation Affects Mesenchymal Stem Cells via Extracellular Oxidized Cell-Free DNA: A Possible Mediator of Bystander Effect and Adaptive Response Oxid Med Cell Longev 2017-09-14 [PMID: 28904740]
• Piechota M, Sunderland P, Wysocka A et al. Is senescence-associated beta-galactosidase a marker of neuronal senescence? Oncotarget. 2016-12-05 [PMID: 27768595] (ICC/IF, Rat)
• Jakl L, Lobachevsky P, Vokalova L et al. Validation of JCountPro software for efficient assessment of ionizing radiation-induced foci in human lymphocytes. Int J Radiat Biol 2016-12-01 [PMID: 27648492] (ICC/IF, Human)
• Wolf C, Rapp A, Berndt N et al. RPA and Rad51 constitute a cell intrinsic mechanism to protect the cytosol from self DNA. Nat Commun. 2016-05-27 [PMID: 27230542] (ICC/IF, Human)
• Onyango DO, Howard SM, Neherin K et al. Tetratricopeptide repeat factor XAB2 mediates the end resection step of homologous recombination. Nucleic Acids Res. 2016-04-15 [PMID: 27084940] (WB, ICC/IF, Human)
• Nassour J, Martien S, Martin N, et al. Defective DNA single-strand break repair is responsible for senescence and neoplastic escape of epithelial cells. Nat Commun. 2016-01-29 [PMID: 26822533] (ICC/IF, WB, Human)
• Markova E, Somsedikova A, Vasilyev S et al. DNA repair foci and late apoptosis/necrosis in peripheral blood lymphocytes of breast cancer patients undergoing radiotherapy. Int. J. Radiat. Biol. 2015-11-04 [PMID: 26537244] (ICC/IF, Human)
• Murakami N, Kuhnel A, Schmid TE et al. Role of membrane Hsp70 in radiation sensitivity of tumor cells. Radiat Oncol 2015-07-22 [PMID: 26197988] (FLOW, Human)
  
  Details:
  Using the Alexa Fluor (R) 488 conjugated version of NB100-78356, catalog number NB100-78356AF488.
• Gunther C, Kind B, Reijns MA et al. Defective removal of ribonucleotides from DNA promotes systemic autoimmunity. J. Clin. Invest. 2014-12-15 [PMID: 25500883] (IHC-P, Human)
• Camps J, Wangsa D, Falke M et al. Loss of lamin B1 results in prolongation of S phase and decondensation of chromosome territories. FASEB J. 2014-04-14 [PMID: 24732130] (ICC/IF, Human)
• Assaily W, Rubinger DA, Wheaton K et al. ROS-mediated p53 induction of Lpin1 regulates fatty acid oxidation in response to nutritional stress. Mol Cell 2011-11-01 [PMID: 22055193]
• Martinelli P et al. Blood. 117:6617. 2011-01-01 [PMID: 21518927]
• Li M et al. Mol Cell Biol. 31:2090. 2011-01-01 [PMID: 21402780]
• Lee J et al. J Cell Biol. 192:263. 2011-01-01 [PMID: 21242291]
• Bu Y et al. Biochem Biophys Res Commun. 397:157. 2010-01-01 [PMID: 20529673] (WB)
• Lukaszewicz A. Chromasoma Apr 27. 2010-01-01 [PMID: 20422424] (ICC/IF)
• Suzuki K, Yamauchi M, Oka Y et al. A novel and simple micro-irradiation technique for creating localized DNA double-strand breaks. Nucleic Acids Res 2010-07-01 [PMID: 20385602] (ICC/IF)
• Yamada C et al. J. Biol. Chem. 285:16693. 2010-01-01 [PMID: 20353948] (WB)
• Massignan T et al. J. Biol Chem. 285:7752. 2010-01-01 [PMID: 19940127] (WB)
• Beels L et al. Circulation. 120:1903. 2009-01-01 [PMID: 19858412] (ICC/IF)
• Loidl J et al. Mol Cell Biol. 20:2048. 2009-01-01 [PMID: 19297526] (ICC/IF)
• Akbay A,et al. Am J Pathol. 173:536. 2008-01-01 [PMID: 18599611] (IF/IHC)
• Moiseenko V et al. Radiat Oncol 3:18. 2008-01-01 [PMID: 18588688] (FLOW)
• Mochizuki K et al. J cell Sci.121:2148. 2008-01-01 [PMID: 18522989] (ICC/IF)
• Rossi DJ et al. Nature. 447:725. 2007-01-01 [PMID: 17554309] (ICC/IF)
• Xiao R et al. Mol Cell Biol.27:5393. 2007-01-01 [PMID: 17526736] (ICC/IF)

Review for gamma H2AX Antibody (NB100-78356) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human and Mouse.

reviewed by: Vincent Picco ICC Human and Mouse 08/18/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: Cultured cell lines
• Species: Human and Mouse
• Lot: B219075

Comments

• Comments: Cell fixed in PBS-2% PFA and permeabilised in PBS-0.1% Triton X100.
  Antibody used 1/300.

Bioinformatics

• Gene Symbol: H2AX