05-636 | Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301

ICC: Jurkat cells were treated with etoposide and stained with Anti-phospho-Histone H2A.X (Ser139), clone JBW301 (green, right panel; DNA stained with DAPI, left panel).
ICC: Jurkat cells were treated with etoposide and stained with Anti-phospho-Histone H2A.X (Ser139),
05-636
200 μg  
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      Overview

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      VrtChIP, ICC, IF, WBMPurifiedMonoclonal Antibody
      Description
      Catalogue Number 05-636
      Brand Family Upstate
      Synonyms Histone H2A.X (phospho S139) H2AXS139P
      Trade Name
      • Upstate
      Description Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301
      Background Information Histone H2A is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N terminal tail H2A is involved with the structure of the nucleosomes of the 'beads on a string' structure.
      References
      Product Information
      Format Purified
      Specificity Recognizes Histone H2A.X phosphorylated at Ser139.
      Control
      • UV-treated 293 cell extracts, UV-treated HeLa cell extracts or breast cancer tissue
      Presentation Immunoaffinity Purified immunoglobulin in 0.1M Tris-Glycine,pH 7.4, 0.15M NaCl, 0.05% sodium azide as a preservative.
      Applications
      Application Use Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301 (Mouse Monoclonal Antibody) validated in ChIP, ICC, IF, WB to detect phospho-Histone H2A.X (Ser139) also known as H2AXS139P.
      Key Applications
      • Chromatin Immunoprecipitation (ChIP)
      • Immunocytochemistry
      • Immunofluorescence
      • Western Blotting
      Application Notes Additional Referenced Applications:
      Chromatin Immunoprecipitation, see Meier, Andreas, et al. EMBO J., 26: 2707-18 (2007) in technical information tab.
      Biological Information
      Immunogen species peptide (C-KATQA[pS]QEY) corresponding to amino acids 134-142 of human histone H2A.X
      Clone JBW301
      Concentration Please refer to the Certificate of Analysis for the lot-specific concentration.
      Host Mouse
      Specificity Recognizes Histone H2A.X phosphorylated at Ser139.
      Isotype IgG1
      Species Reactivity Vertebrates
      Antibody Type Monoclonal Antibody
      Entrez Gene Number
      Entrez Gene Summary Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif.
      Gene Symbol
      • H2A.X
      • H2AFX
      • H2AX
      • H2a/x
      • H2A/X
      Modifications
      • Phosphorylation
      Purification Method Protein G purfied
      UniProt Number
      UniProt Summary FUNCTION: SwissProt: P16104 # Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C- terminal phosphorylation.
      SIZE: 143 amino acids; 15145 Da
      SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with numerous proteins required for DNA damage signaling and repair when phosphorylated on Ser-140. These include MDC1, TP53BP1, BRCA1 and the MRN complex, composed of MRE11A, RAD50, and NBN. Interaction with the MRN complex is mediated at least in part by NBN. Also interacts with DHX9/NDHII when phosphorylated on Ser-140.
      SUBCELLULAR LOCATION: Nucleus.DEVELOPMENTAL STAGE: Synthesized in G1 as well as in S-phase.
      DOMAIN: SwissProt: P16104 The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family.
      PTM: Phosphorylated on Ser-140 (to form gamma-H2AFX) in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Phosphorylation of Ser-140 in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation may also be required for transcriptional repression of unsynapsed chromatin and meiotic sex chromosome inactivation (MSCI), whereby the X and Y chromosomes condense in pachytene to form the heterochromatic XY-body. During immunoglobulin class switch recombination in lymphocytes, Ser-140 phosphorylation may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. & Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression (By similarity).
      SIMILARITY: Belongs to the histone H2A family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assurance Immunoblot Analysis: 0.05-1 μg/ml of this antibody detected phosphorylated histone H2A.X (Ser139) in acid extracted histone lysates from Jurkat cells treated with 0.5 μM staurosporine (Catalog # 19-123).
      Immunocytochemistry: 2 μg/ml of this antibody detected phosphorylated histone H2A.X in HeLa cells treated with 0.5 μM staurosporine for 4-6 hours.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage Conditions Maintain for 1 year at 2 to 8°C from date of shipment. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
      Packaging Information
      Material Size 200 μg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

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      Certificates of Analysis

      TitleLot Number
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 2476967
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 (mouse monoclonal IgG1) - DAM1405597 DAM1405597
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 (mouse monoclonal IgG1) - DAM1474315 DAM1474315
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2116620 2116620
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2390526 2390526
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2392265 2392265
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2420445 2420445
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2424700 2424700
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2428660 2428660
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2441914 2441914
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2455602 2455602
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 0702052447 0702052447
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 1997719 1997719
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2019732 2019732
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2064512 2064512
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2068177 2068177
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2138016 2138016
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2202540 2202540
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 22092 22092
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2211083 2211083
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2227256 2227256
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2239793 2239793
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2250524 2250524
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2275586 2275586
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2276332 2276332
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2310355 2310355
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 23760 23760
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 2506483 2506483
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 25358 25358
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 27505 27505
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - 32526 32526
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1493341 DAM1493341
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1567248 DAM1567248
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1636071 DAM1636071
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1661039 DAM1661039
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1690636 DAM1690636
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1772418 DAM1772418
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - DAM1782241 DAM1782241
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - JBC1353261 JBC1353261
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - JBC1367868 JBC1367868
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - JBC1863420 JBC1863420
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - JBC1881392 JBC1881392
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - NG1904671 NG1904671
      Anti-phospho-Histone H2A.X (Ser139), clone JBW301 - NG1951659 NG1951659

      References | 98 Available | See All References

      Reference overviewPub Med ID
      PARP-1 dependent recruitment of the amyotrophic lateral sclerosis-associated protein FUS/TLS to sites of oxidative DNA damage.
      Rulten, Stuart L, et al.
      Nucleic Acids Res., 42: 307-14 (2014) 2014

      Show Abstract
      24049082
      APLF promotes the assembly and activity of non-homologous end joining protein complexes.
      Grundy, Gabrielle J, et al.
      EMBO J., 32: 112-25 (2013) 2013

      Show Abstract
      23178593
      Acetylation limits 53BP1 association with damaged chromatin to promote homologous recombination.
      Tang, Jiangbo, et al.
      Nat. Struct. Mol. Biol., 20: 317-25 (2013) 2013

      Show Abstract
      23377543
      Homologous recombination DNA repair genes play a critical role in reprogramming to a pluripotent state.
      González, Federico, et al.
      Cell Rep, 3: 651-60 (2013) 2013

      Show Abstract
      23478019
      PARP inhibition sensitizes to low dose-rate radiation TMPRSS2-ERG fusion gene-expressing and PTEN-deficient prostate cancer cells.
      Chatterjee, Payel, et al.
      PLoS ONE, 8: e60408 (2013) 2013

      Show Abstract
      23565244
      Loss of WSTF results in spontaneous fluctuations of heterochromatin formation and resolution, combined with substantial changes to gene expression.
      Culver-Cochran, Ashley E and Chadwick, Brian P
      BMC Genomics, 14: 740 (2013) 2013

      Show Abstract
      24168170
      The activation of DNA damage detection and repair responses in cleavage-stage rat embryos by a damaged paternal genome.
      Lisanne Grenier,Bernard Robaire,Barbara F Hales
      Toxicological sciences : an official journal of the Society of Toxicology 127 2012

      Show Abstract
      22454429
      Role of SUMO modification of human PCNA at stalled replication fork.
      Himabindu Gali,Szilvia Juhasz,Monika Morocz,Ildiko Hajdu,Karoly Fatyol,Valeria Szukacsov,Peter Burkovics,Lajos Haracska
      Nucleic acids research 40 2012

      Show Abstract
      22457066
      The p21-dependent radiosensitization of human breast cancer cells by MLN4924, an investigational inhibitor of NEDD8 activating enzyme.
      Yang, Dong, et al.
      PLoS ONE, 7: e34079 (2012) 2012

      Show Abstract
      22457814
      The autophagy-senescence connection in chemotherapy: must tumor cells (self) eat before they sleep?
      Rachel W Goehe,Xu Di,Khushboo Sharma,Molly L Bristol,Scott C Henderson,Kristoffer Valerie,Francis Rodier,Albert R Davalos,David A Gewirtz
      The Journal of pharmacology and experimental therapeutics 343 2012

      Show Abstract
      22927544

      Brochure

      Title
      Advancing cancer research: From hallmarks & biomarkers to tumor microenvironment progression

      FAQ

      QuestionAnswer
      How can I check the phosphospecificity of my antibody once my protein is already immobilized on a membrane?You can check for the phosphospecificity of your antibody using Lambda Protein Phosphatase. Simply perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell lysate and transfer the proteins to your membrane of choice. Wash the blotted nitrocellulose twice with water. Block the blotted nitrocellulose in freshly prepared TBS containing 1% bovine serum albumin (BSA) and 0.1% Triton X-100 for 1 hour at 20-25°C with constant agitation. Incubate the nitrocellulose in TBS containing 1% bovine serum albumin (BSA), 0.1% Triton X-100 and 2 mM MnCl2, and where dephosphorylation of proteins is desirable, 400 U/ml Lambda Protein Phosphatase for two hours at room temperature, or overnight at 4°C. After incubation, wash the nitrocellulose in PBS-0.1% Tween 20 for 3-5 minutes. Rinse the nitrocellulose in 4-5 changes of water. Continue with your western blotting assay.

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      Alternative Packsize

       Catalogue Number Description  
      05-636-KC Anti-Phos-Hist H2A.1 (S139) (KC) Show Pricing & Availability

      Alternative Format

       Catalogue Number Description  
      16-193 Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, biotin conjugate Show Pricing & Availability
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