Histone H2AX [p Ser139] Antibody [Unconjugated]

Histone H2AX was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 μM camptothecin (CPT) for for the indicated times. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Histone H2AX (S139) at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 µM Camptothecin (CPT) for 24 hours, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Histone H2AX (S139) at approximately 25 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Histone H2AX phosphorylated at S139 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with ultraviolet radiation using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

Histone H2AX [p Ser139] Antibody [Unconjugated] Summary

• Immunogen: Phosphopeptide containing human Histone H2AX S139 site
• Modification: p Ser139
• Specificity: Detects human Histone H2AX when phosphorylated at S139.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: H2AFX
• Purity: Antigen and protein A Affinity-purified
• Purity Statement: Antigen and protein A Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 5 ug/mL
  • Western Blot 0.5 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Antigen and protein A Affinity-purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Histone H2AX [p Ser139] Antibody [Unconjugated]

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

Background

Histone H2AX is phosphorylated at S139 in cells exposed to DNA double-strand break-inducing agents, such as ionizing radiation. The S139 phosphorylated H2AX, termed ( gamma -H2AX, marks the site of DNA double-strand breaks and serves to recruit cell cycle checkpoint and DNA repair factors to the site of damage.

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 (AF2288)(18)

Publications using AF2288

• Nataliya, B;Mikhail, A;Vladimir, P;Olga, G;Maksim, V;Ivan, Z;Ekaterina, N;Georgy, S;Natalia, D;Pavel, M;Andrey, C;Maria, S;Maxim, K;Anastasiya, T;Uliana, D;Zhanna, A;Vsevolod, T;Natalia, K;Anastasiya, E; Mesenchymal stromal cells facilitate resolution of pulmonary fibrosis by miR-29c and miR-129 intercellular transfer Experimental & molecular medicine 2023-07-03 [PMID: 37394579] (Western Blot, Human)
• DM Zhang, R Navara, T Yin, J Szymanski, U Goldsztejn, C Kenkel, A Lang, C Mpoy, CE Lipovsky, Y Qiao, S Hicks, G Li, KMS Moore, C Bergom, BE Rogers, CG Robinson, PS Cuculich, JK Schwarz, SL Rentschler Cardiac radiotherapy induces electrical conduction reprogramming in the absence of transmural fibrosis Nature Communications, 2021-09-24;12(1):5558. 2021-09-24 [PMID: 34561429] (IHC, Mouse)
• M Al-Mugotir, JJ Lovelace, J George, M Bessho, D Pal, L Struble, C Kolar, S Rana, A Natarajan, T Bessho, GEO Borgstahl Selective killing of homologous recombination-deficient cancer cell lines by inhibitors of the RPA:RAD52 protein-protein interaction PLoS ONE, 2021-03-30;16(3):e0248941. 2021-03-30 [PMID: 33784323] (Western Blot, Human)
• A Schepsky, G Asta Traus, J Petur Joel, S Ingthorsso, J Kricker, J Thor Bergt, A Asbjarnars, T Gudjonsson, N Nupponen, A Slipicevic, F Lehmann, T Gudjonsson Melflufen, a peptide-conjugated alkylator, is an efficient anti-neo-plastic drug in breast cancer cell lines Cancer Med, 2020-07-27;0(0):. 2020-07-27 [PMID: 32717133] (ICC, Human)
• CE Hellweg, V Shinde, SP Srinivasan, M Henry, T Rotshteyn, C Baumstark-, C Schmitz, S Feles, LF Spitta, R Hemmersbac, J Hescheler, A Sachinidis Radiation Response of Murine Embryonic Stem Cells Cells, 2020-07-09;9(7):. 2020-07-09 [PMID: 32660081] (ICC, Mouse)
• S Miranda, M Correia, AG Dias, A Pestana, P Soares, J Nunes, J Lima, V Máximo, P Boaventura Evaluation of the role of mitochondria in the non-targeted effects of ionizing radiation using cybrid cellular models Sci Rep, 2020-04-09;10(1):6131. 2020-04-09 [PMID: 32273537] (ICC, Human)
• HL Li, CY Wang, J Fu, XJ Yang, Y Sun, YH Shao, LH Zhang, XM Yang, XL Zhang, J Lin PTEN expression in U251 glioma cells enhances their sensitivity to ionizing radiation by suppressing DNA repair capacity Eur Rev Med Pharmacol Sci, 2019-12-01;23(23):10453-10458. 2019-12-01 [PMID: 31841199] (ICC, Human)
• Y Wang, AJ Bernhardy, J Nacson, JJ Krais, YF Tan, E Nicolas, MR Radke, E Handorf, A Llop-Gueva, J Balmaña, EM Swisher, V Serra, S Peri, N Johnson BRCA1 intronic Alu elements drive gene rearrangements and PARP inhibitor resistance Nat Commun, 2019-12-11;10(1):5661. 2019-12-11 [PMID: 31827092] (ICC, Human)
• Z Simandi, K Pajer, K Karolyi, T Sieler, LL Jiang, Z Kolostyak, Z Sari, Z Fekecs, A Pap, A Patsalos, GA Contreras, B Reho, Z Papp, X Guo, A Horvath, G Kiss, Z Keresztess, G Vámosi, J Hickman, H Xu, D Dormann, T Hortobagyi, M Antal, A Nogradi, L Nagy Arginine methyltransferase PRMT8 provides cellular stress tolerance in aging motoneurons J. Neurosci., 2018-07-27;0(0):. 2018-07-27 [PMID: 30054395] (IHC, Western Blot, Mouse)
• M Jangi, C Fleet, P Cullen, SV Gupta, S Mekhoubad, E Chiao, N Allaire, CF Bennett, F Rigo, AR Krainer, JA Hurt, JP Carulli, JF Staropoli SMN deficiency in severe models of spinal muscular atrophy causes widespread intron retention and DNA damage Proc. Natl. Acad. Sci. U.S.A, 2017-03-07;0(0):. 2017-03-07 [PMID: 28270613] (IHC, Mouse)
• N Rebolleda, I Losada-Fer, G Perez-Chac, R Castejon, S Rosado, M Morado, MT Vallejo-Cr, A Martinez, JA Vargas-Nuñ, P Perez-Acie Synergistic Activity of Deguelin and Fludarabine in Cells from Chronic Lymphocytic Leukemia Patients and in the New Zealand Black Murine Model PLoS ONE, 2016-04-21;11(4):e0154159. 2016-04-21 [PMID: 27101369] (Western Blot, Human)
• Garwood C, Ratcliffe L, Morgan S, Simpson J, Owens H, Vazquez-Villasenor I, Heath P, Romero I, Ince P, Wharton S Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo; characterisation, subcellular localisation and modulation of the receptors. Mol Brain, 2015-08-22;8(0):51. 2015-08-22 [PMID: 26297026]
• Kim , Bo S, Serebreni , Leonid, Fallica , Jonathan, Hamdan , Omar, Wang , Lan, Johnston , Laura, Kolb , Todd, Damarla , Mahendra, Damico , Rachel, Hassoun , Paul M Cyclin-dependent kinase five mediates activation of lung xanthine oxidoreductase in response to hypoxia. PLoS ONE, 2015-04-01;10(4):e0124189. 2015-04-01 [PMID: 25831123] (Western Blot, Rat)
• Jia XZ, Yang SY, Zhou J, Li SY, Ni JH, An GS, Jia HT Inhibition of CHK1 kinase by Go6976 converts 8-chloro-adenosine-induced G2/M arrest into S arrest in human myelocytic leukemia K562 cells. Biochem. Pharmacol., 2008-11-18;77(5):770-80. 2008-11-18 [PMID: 19059218] (ICC, Western Blot, Human)
• Yang SY, Jia XZ, Feng LY, Li SY, An GS, Ni JH, Jia HT Inhibition of topoisomerase II by 8-chloro-adenosine triphosphate induces DNA double-stranded breaks in 8-chloro-adenosine-exposed human myelocytic leukemia K562 cells. Biochem. Pharmacol., 2008-10-28;77(3):433-43. 2008-10-28 [PMID: 19014910] (ICC, Western Blot, Human)
• Roh DS, Cook AL, Rhee SS, Joshi A, Kowalski R, Dhaliwal DK, Funderburgh JL DNA cross-linking, double-strand breaks, and apoptosis in corneal endothelial cells after a single exposure to mitomycin C. Invest. Ophthalmol. Vis. Sci., 2008-07-24;49(11):4837-43. 2008-07-24 [PMID: 18658091] (ICC, Goat)
• Tang KF, He CX, Zeng GL, Wu J, Song GB, Shi YS, Zhang WG, Huang AL, Steinle A, Ren H Induction of MHC class I-related chain B (MICB) by 5-aza-2'-deoxycytidine. Biochem. Biophys. Res. Commun., 2008-04-04;370(4):578-83. 2008-04-04 [PMID: 18395517] (ICC, Human)
• Bogliolo M, Lyakhovich A, Callen E, Castella M, Cappelli E, Ramirez MJ, Creus A, Marcos R, Kalb R, Neveling K, Schindler D, Surralles J Histone H2AX and Fanconi anemia FANCD2 function in the same pathway to maintain chromosome stability. EMBO J., 2007-02-15;26(5):1340-51. 2007-02-15 [PMID: 17304220] (ICC, Human)

Bioinformatics

• Gene Symbol: H2AFX