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

05-636
200 µg  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      VrtICC, IF, WB, ChIPMPurifiedMonoclonal Antibody
      Description
      Catalogue Number05-636
      ReplacesMABE205
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionAnti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301
      Alternate Names
      • H2AXS139P
      • Histone H2A.X (phospho S139)
      Background InformationHistone 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
      FormatPurified
      Control
      • UV-treated 293 cell extracts, UV-treated HeLa cell extracts or breast cancer tissue
      PresentationImmunoaffinity Purified immunoglobulin in 0.1M Tris-Glycine,pH 7.4, 0.15M NaCl, 0.05% sodium azide as a preservative.
      Applications
      ApplicationAnti-phospho-Histone H2A.X (Ser139), clone JBW301 is a well published Mouse Monoclonal Antibody validated in ChIP, ICC, IF, WB. This purified mAb is highly specific for phospho-Histone H2A.X (Ser139) also known as H2AXS139p.
      Key Applications
      • Immunocytochemistry
      • Immunofluorescence
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesAdditional Referenced Applications:
      Chromatin Immunoprecipitation, see Meier, Andreas, et al. EMBO J., 26: 2707-18 (2007) in technical information tab.
      Biological Information
      Immunogenpeptide (C-KATQA[pS]QEY) corresponding to amino acids 134-142 of human histone H2A.X
      CloneJBW301
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityRecognizes Histone H2A.X phosphorylated at Ser139.
      IsotypeIgG1
      Species Reactivity
      • Vertebrates
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones 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
      • H2AFX
      • H2AX
      • H2a/x
      • H2A/X
      • H2A.X
      Modifications
      • Phosphorylation
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFUNCTION: 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.
      Molecular Weight17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceImmunoblot 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 ConditionsMaintain 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 Size200 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-phospho-Histone H2A.X (Ser139), -2739172 2739172
      Anti-phospho-Histone H2A.X (Ser139), -2789757 2789757
      Anti-phospho-Histone H2A.X (Ser139), -2806127 2806127
      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

      References

      Reference overviewApplicationSpeciesPub Med ID
      In vivo measurement of dose distribution in patients' lymphocytes: helical tomotherapy versus step-and-shoot IMRT in prostate cancer.
      Zwicker, F; Swartman, B; Roeder, F; Sterzing, F; Hauswald, H; Thieke, C; Weber, KJ; Huber, PE; Schubert, K; Debus, J; Herfarth, K
      Journal of radiation research  56  239-47  2015

      Show Abstract
      25361548 25361548
      Co-visualization of DNA damage and ion traversals in live mammalian cells using a fluorescent nuclear track detector.
      Kodaira, S; Konishi, T; Kobayashi, A; Maeda, T; Ahmad, TA; Yang, G; Akselrod, MS; Furusawa, Y; Uchihori, Y
      Journal of radiation research  56  360-5  2015

      Show Abstract
      25324538 25324538
      TIMELESS Forms a Complex with PARP1 Distinct from Its Complex with TIPIN and Plays a Role in the DNA Damage Response.
      Young, LM; Marzio, A; Perez-Duran, P; Reid, DA; Meredith, DN; Roberti, D; Star, A; Rothenberg, E; Ueberheide, B; Pagano, M
      Cell reports  13  451-9  2015

      Show Abstract
      26456830 26456830
      HMGB1 facilitates repair of mitochondrial DNA damage and extends the lifespan of mutant ataxin-1 knock-in mice.
      Ito, H; Fujita, K; Tagawa, K; Chen, X; Homma, H; Sasabe, T; Shimizu, J; Shimizu, S; Tamura, T; Muramatsu, S; Okazawa, H
      EMBO molecular medicine  7  78-101  2015

      Show Abstract
      25510912 25510912
      SETD2 loss-of-function promotes renal cancer branched evolution through replication stress and impaired DNA repair.
      Kanu, N; Grönroos, E; Martinez, P; Burrell, RA; Yi Goh, X; Bartkova, J; Maya-Mendoza, A; Mistrík, M; Rowan, AJ; Patel, H; Rabinowitz, A; East, P; Wilson, G; Santos, CR; McGranahan, N; Gulati, S; Gerlinger, M; Birkbak, NJ; Joshi, T; Alexandrov, LB; Stratton, MR; Powles, T; Matthews, N; Bates, PA; Stewart, A; Szallasi, Z; Larkin, J; Bartek, J; Swanton, C
      Oncogene  34  5699-708  2015

      Show Abstract
      25728682 25728682
      Ablation of the p16(INK4a) tumour suppressor reverses ageing phenotypes of klotho mice.
      Sato, S; Kawamata, Y; Takahashi, A; Imai, Y; Hanyu, A; Okuma, A; Takasugi, M; Yamakoshi, K; Sorimachi, H; Kanda, H; Ishikawa, Y; Sone, S; Nishioka, Y; Ohtani, N; Hara, E
      Nature communications  6  7035  2015

      Show Abstract
      25923845 25923845
      Resveratrol Induced Premature Senescence Is Associated with DNA Damage Mediated SIRT1 and SIRT2 Down-Regulation.
      Kilic Eren, M; Kilincli, A; Eren, Ö
      PloS one  10  e0124837  2015

      Show Abstract
      25924011 25924011
      Dexamethasone acts as a radiosensitizer in three astrocytoma cell lines via oxidative stress.
      Ortega-Martínez, S
      Redox biology  5  388-97  2015

      Show Abstract
      26160768 26160768
      DNA ligase III acts as a DNA strand break sensor in the cellular orchestration of DNA strand break repair.
      Abdou, I; Poirier, GG; Hendzel, MJ; Weinfeld, M
      Nucleic acids research  43  875-92  2015

      Show Abstract
      25539916 25539916
      SWI/SNF complexes are required for full activation of the DNA-damage response.
      Smith-Roe, SL; Nakamura, J; Holley, D; Chastain, PD; Rosson, GB; Simpson, DA; Ridpath, JR; Kaufman, DG; Kaufmann, WK; Bultman, SJ
      Oncotarget  6  732-45  2015

      Show Abstract
      Western Blotting25544751 25544751

      Brochure

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

      Technical Info

      Title
      White Paper - The Message in the Marks: Deciphering Cancer Epigenetics

      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 Format

      Catalogue Number Description  
      16-193 Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, biotin conjugate Show Pricing & Availability
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      Life Science Research > Antibodies and Assays > Primary Antibodies