Overview

  • Product name

    Anti-mH2A1 antibody [EPR9359(2)]
    See all mH2A1 primary antibodies
  • Description

    Rabbit monoclonal [EPR9359(2)] to mH2A1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human mH2A1 aa 200-300. The exact sequence is proprietary.
    Database link: O75367

  • Positive control

    • WB: HepG2, MCF7, 293T and HeLa whole cell lysate (ab150035) IHC-P: Human kidney and liver tissues ICC-IF: HAP1-WT and H2AFY knockout cells. MCF7 and HeLa cells.
  • General notes

    Previously labelled as macroH2A.1.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR9359(2)
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183041 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 1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
IHC-P 1/50 - 1/100.
ICC/IF Use a concentration of 1 µg/ml.

This antibody gives positive signal in both 4%PFA and 100% MeOH-fixed cells.

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

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: mH2A1 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 - ab183041 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab183041 was shown to specifically react with mH2A1 when mH2A1 knockout samples were used. Wild-type and mH2A1 knockout samples were subjected to SDS-PAGE. Ab183041 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

     

     

  • ab183041 staining mH2A1 in HAP1 WT and H2AFY knockout cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1%PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab183041 at 1μg/ml and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594) at 1/250 dilution (shown in pseudocolor red). Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver from wild-type and mH2A1/2 knock out tissue sections labeling mH2A1 with ab183041 at 1/400 dilution. Sections were fixed in formaldehyde; heat mediated antigen retivial was performed using a citrate buffer pH 6. An undiluted polyclonal horse anti-rabbit IgG (HRP-conjugated) was used as the secondary antibody.

    See Abreview

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed MCF7 cells labeling mH2A1 with ab183041 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 488) at 1/200 dilution (green). Counter stained with Dapi (blue).

  • All lanes : Anti-mH2A1 antibody [EPR9359(2)] (ab183041) at 1/50000 dilution

    Lane 1 : HepG2 cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa

  • Immunofluorescent analysis of acetone-fixed HeLa cells labeling mH2A1 with ab183041 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 488) at 1/200 dilution (green). Counter stained with Dapi (blue).

  • Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling mH2A1 with ab183041 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling mH2A1 with ab183041 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

References

This product has been referenced in:

  • Baklaushev VP  et al. Tissue Engineered Neural Constructs Composed of Neural Precursor Cells, Recombinant Spidroin and PRP for Neural Tissue Regeneration. Sci Rep 9:3161 (2019). Read more (PubMed: 30816182) »
  • Li W  et al. Genome-Wide RNAi Screen Identify Melanoma-Associated Antigen Mageb3 Involved in X Chromosome Inactivation. J Mol Biol N/A:N/A (2018). ICC/IF . Read more (PubMed: 29800566) »
See all 5 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver from WT and macroH2A1/2 KO)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Specification
Liver from WT and macroH2A1/2 KO
Blocking step
PBS, 2% horse serum, 1% BSA, 0.1% Triton X-100, 0.05% Tween 20, 0.05% sodium azide as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Formaldehyde

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Submitted Nov 16 2016

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