Overview

  • Product name
    Anti-macroH2A.1 antibody [14G7]
    See all macroH2A.1 primary antibodies
  • Description
    Mouse monoclonal [14G7] to macroH2A.1
  • Tested applications
    Suitable for: WB, ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein of Human macroH2A.1.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; MCF7
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity
    IgG fraction
  • Clonality
    Monoclonal
  • Clone number
    14G7
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab91528 in the following tested applications.

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

Application Abreviews Notes
WB Use a concentration of 10 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
ELISA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Variant histone H2A which replaces conventional H2A in a subset of nucleosomes where it represses transcription. 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. Involved in stable X chromosome inactivation. Inhibits the binding of transcription factors and interferes with the activity of remodeling SWI/SNF complexes. Inhibits histone acetylation by EP300 and recruits class I HDACs, which induces an hypoacetylated state of chromatin. In addition, isoform 1, but not isoform 2, binds ADP-ribose and O-acetyl-ADP-ribose, and may be involved in ADP-ribose-mediated chromatin modulation.
  • Tissue specificity
    Ubiquitous.
  • Sequence similarities
    Contains 1 histone H2A domain.
    Contains 1 Macro domain.
  • Post-translational
    modifications
    Monoubiquitinated at either Lys-116 or Lys-117. May also be polyubiquitinated. Ubiquitination is mediated by the CUL3/SPOP E3 complex and does not promote proteasomal degradation. Instead, it is required for enrichment in inactive X chromosome chromatin.
  • Cellular localization
    Nucleus. Chromosome. Enriched in inactive X chromosome chromatin and in senescence-associated heterochromatin.
  • Information by UniProt
  • Database links
  • Alternative names
    • Core histone macro h2a.1 antibody
    • Core histone macro-H2A.1 antibody
    • H2A histone family member Y antibody
    • H2A.y antibody
    • H2A/y antibody
    • H2AF12M antibody
    • H2AFJ antibody
    • H2afy antibody
    • H2AY_HUMAN antibody
    • Histone H2A.Y antibody
    • Histone macroH2A1 antibody
    • Histone macroH2A1.1 antibody
    • Histone macroH2A1.2 antibody
    • Macroh2a1 antibody
    • MACROH2A1.1 antibody
    • MacroH2A1.2 antibody
    • Medulloblastoma antigen MU MB 50.205 antibody
    • Medulloblastoma antigen MU-MB-50.205 antibody
    • mH2a antibody
    • mH2A1 antibody
    see all

Images



  • Predicted band size : 40 kDa

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: MacroH2A.1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HepG2 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab91528 observed at 40 kDa. Red - loading control, ab176560, observed at 50 kDa.

    ab91528 was shown to specifically recognize MacroH2A.1 in wild-type HAP1 cells. No band was observed when MacroH2A.1 knockout cells were examined. Wild-type and MacroH2A.1 knockout samples were subjected to SDS-PAGE. ab91528 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 10 µg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Overlay histogram showing HeLa cells stained with ab91528 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab91528, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-macroH2A.1 antibody [14G7] (ab91528) at 10 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa
    Additional bands at : 14 kDa,18 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

References

ab91528 has not yet been referenced specifically in any publications.

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

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