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Overview

  • Product nameAnti-Histone H2A.X antibodySee all Histone H2A.X primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H2A.X
  • Tested applicationsWB, ELISA more details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Cow, Dog, Pig, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to a region within N terminal amino acids 2-51 (SGRGKTGGKA RAKAKSRSSR AGLQFPVGRV HRLLRKGHYA ERVGAGAPVY) of Human Histone H2A.X (NP_002096).

  • Positive controlHepG2 cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab83787 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB
ELISA
  • Application notesELISA titre using peptide based assay: 1/312500.
    WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionVariant 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.
    • Sequence similaritiesBelongs to the histone H2A family.
    • Developmental stageSynthesized in G1 as well as in S-phase.
    • DomainThe [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family.
    • Post-translational
      modifications
      Phosphorylated on Ser-140 (to form gamma-H2AFX or H2AX139ph) 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 (H2AX139ph) in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation (H2AX139ph) subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation (H2AX139ph) may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation (H2AX139ph) may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation (H2AX139ph) 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 (H2AX139ph) may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. Phosphorylation at Tyr-143 (H2AXY142ph) by BAZ1B/WSTF determines the relative recruitment of either DNA repair or pro-apoptotic factors. Phosphorylation at Tyr-143 (H2AXY142ph) favors the recruitment of APBB1/FE65 and pro-apoptosis factors such as MAPK8/JNK1, triggering apoptosis. In contrast, dephosphorylation of Tyr-143 by EYA proteins (EYA1, EYA2, EYA3 or EYA4) favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated Ser-140 (H2AX139ph).
      Monoubiquitination of Lys-120 (H2AXK119ub) by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events.
    • Cellular localizationNucleus. Chromosome.
    • Target information above from: UniProt accession P16104 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links
    • Alternative names
        AW228881 antibodyH2A histone family member X antibodyH2A.FX antibody
        H2A.X antibodyH2A/X antibodyH2AFX antibodyH2AX antibodyH2AX histone antibodyH2AX_HUMAN antibodyHist5.2ax antibodyHistone 2A antibodyHistone 2AX antibodyHistone H2A.x antibodyRGD1566119 antibody
      see all

    Anti-Histone H2A.X antibody images

    • Anti-Histone H2A.X antibody (ab83787) at 1 µg/ml (5% skim milk / PBS buffer) + HepG2 cell lysate at 10 µg

      Secondary
      HRP conjugated anti-Rabbit IgG at 1/50000 dilution

      Predicted band size : 15 kDa
      Observed band size : 15 kDa

    References for Anti-Histone H2A.X antibody (ab83787)

    This product has been referenced in:

    See 1 Publication for this product

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    Application Western blot
    Sample Chicken Purified protein (acid extracted histones)
    Loading amount 100 µg
    Specification acid extracted histones
    Gel Running Conditions Reduced Denaturing (15%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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    Submitted Nov 19 2010

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"