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).

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
• Applications: WB, Simple Western, ELISA, Flow, ICC/IF, IHC
• Clone: 3F2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Histone H2AX [p Ser139] Antibody (3F2) Summary

• Immunogen: This Histone H2AX [p Ser139] Antibody (3F2) was developed against a synthetic peptide sequence surrounding phosphorylated Ser139.
• 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.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: H2AX
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • 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.
• 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.
• Control:

  Histone H2AX Overexpression Lysate

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.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS with 1 mg/ml BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein 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 (3F2)

• 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 Histone H2AX Antibody (NB100-74435)(18)

Publications using NB100-74435

• Singh M, Kondrashkina AM, Widmann TJ et al. A new human embryonic cell type associated with activity of young transposable elements allows definition of the inner cell mass PLoS biology 2023-06-01 [PMID: 37339119]
• Trier I, Black EM, Joo YK, Kabeche L ATR protects centromere identity by promoting DAXX association with PML nuclear bodies Cell reports 2023-05-30 [PMID: 37163376] (ICC/IF, Human)
• Vigasova K, Durdik M, Jakl L et al. Chemotherapy and cryopreservation affects DNA repair foci in lymphocytes of breast cancer patients International journal of radiation biology 2023-05-18 [PMID: 37145321]
• Bhardwaj P, Iyengar NM, Zahid H et al. Obesity promotes breast epithelium DNA damage in women carrying a germline mutation in BRCA1 or BRCA2 Science translational medicine 2023-02-22 [PMID: 36812344]
• Admasu TD, Kim K, Rae M et al. Selective ablation of primary and paracrine senescent cells by targeting iron dyshomeostasis Cell reports 2023-02-06 [PMID: 36753419] (ICC/IF, Human)
• Kim K, Admasu TD, Stolzing A, Sharma A Enhanced co-culture and enrichment of human natural killer cells for the selective clearance of senescent cells Aging 2022-03-04 [PMID: 35245208] (ICC/IF, Human)
• PAramo M, SantamarIa E, Idoate MA et al. A new animal model of atrophy-hypertrophy complex and liver damage following Yttrium-90 lobar selective internal radiation therapy in rabbits Scientific reports 2022-02-02 [PMID: 35110610] (IHC-P, Rabbit)
• Sugaya K Characterization of cells expressing MRI reporters for the analysis of epigenetics Analytical biochemistry 2021-09-30 [PMID: 34600867] (IM)
• Wang H, Muthu Karuppan M, Devadoss D et al. TFEB protein expression is reduced in aged brains and its overexpression mitigates senescence-associated biomarkers and memory deficits in mice Neurobiology of Aging 2021-10-01 [PMID: 34229273]
• Bui TM, Butin-Israeli V, Wiesolek HL et al. Neutrophils Alter DNA Repair Landscape to Impact Survival and Shape Distinct Therapeutic Phenotypes of Colorectal Cancer Gastroenterology 2021-03-19 [PMID: 33753103] (Mouse)
• Kobayashi, Y, Tata, A Et al. Persistence of a regeneration-associated, transitional alveolar epithelial cell state in pulmonary fibrosis. Nat Cell Biol 2020-08-01 [PMID: 32661339] (IP, Human)
• Elsaid MY, Shahi A, Wang AR et al. Enhanced Radiosensitivity in Solid Tumors using a Tumor-Selective Alkyl Phosphocholine Analog Mol. Cancer Ther. 2018-08-14 [PMID: 30108133] (IHC-P, Human)
• Hayashi M, Yoshida K, Kitada K et al. Low-dose irradiation of mouse embryos increases Smad-p21 pathway activity and preserves pluripotency J. Assist. Reprod. Genet. 2018-03-16 [PMID: 29546598] (WB, Mouse)
• Szczepny A, Carey K, McKenzie L et al. The tumor suppressor Hic1 maintains chromosomal stability independent of Tp53 Oncogene 2018-01-25 [PMID: 29367758] (WB, Mouse)
• Pelz L, Purfurst B, et al. The cell adhesion molecule BT-IgSF is essential for a functional blood-testis barrier and male fertility in mice. J Biol Chem 2017-12-29 [PMID: 29123028] (IF/IHC, Mouse)
• Hatanaka M, Anderson-Baucum E, Lakhter A et al. Chronic high fat feeding restricts islet mRNA translation initiation independently of ER stress via DNA damage and p53 activation Sci Rep. 2017-06-18 [PMID: 28630491] (IHC-P, Human)
• Singh M, Widmann T, Cortes J et al. The phylogenetically distinct early human embryo bioRxiv 2018-05-18 (IF/IHC, Human)
• Bui TM Dissecting the Diverse Phenotypes and Pathological Impacts of Neutrophils in Colitis-to-CRC Progression Thesis 2022-01-01

Review for Histone H2AX Antibody (NB100-74435) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer IF Human 05/17/2022

Summary

• Application: Immunofluorescence
• Sample Tested: MDA MB 231 cells
• Species: Human
• Lot: VL317613

Bioinformatics

• Gene Symbol: H2AX
• Entrez:
  • 3014(Human)
  • 15270(Mouse)
• Uniprot:
  • P16104()