Anti-macroH2A.1 antibody - ChIP Grade (ab37264)

Overview

  • Product name
    Anti-macroH2A.1 antibody - ChIP Grade
    See all macroH2A.1 primary antibodies
  • Description
    Rabbit polyclonal to macroH2A.1 - ChIP Grade
  • Specificity
    We have had varying reports about the efficiency with which this antibody recognises macroH2A.1 in mouse cells and tissues. Please contact our Scientific Support team if you have any queries about this.
  • Tested applications
    Suitable for: ChIP, Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Cow, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human macroH2A.1.

    (Peptide available as ab37263.)

  • Positive control
    • This antibody gave a positive signal in Hela, HEK293 and NIH3T3 cell lysates

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab37264 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
ChIP Use at an assay dependent concentration. PubMed: 19380460
Flow Cyt Use at an assay dependent concentration. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).Can be blocked with Rat macroH2A.1 peptide (ab37263).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 - 5 µg/ml.

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 - ab37264 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab37264 was shown to specifically recognize macroH2A.1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when macroH2A.1 knockout cells were examined. Wild-type and macroH2A.1 knockout samples were subjected to SDS-PAGE. Ab37264 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • ab37264 stained in Hela cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab37264 at 5 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 µg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • IHC image of macroH2A.1 staining in human skin FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab37264, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-macroH2A.1 antibody - ChIP Grade (ab37264) at 1 µg

    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 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa
    Additional bands at : 100 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 16 minutes

References

This product has been referenced in:
  • Park SJ  et al. MacroH2A1 downregulation enhances the stem-like properties of bladder cancer cells by transactivation of Lin28B. Oncogene 35:1292-301 (2016). WB, IHC ; Human . Read more (PubMed: 26028027) »
  • Sinha VC  et al. A p53/ARF-dependent anticancer barrier activates senescence and blocks tumorigenesis without impacting apoptosis. Mol Cancer Res 13:231-8 (2015). Read more (PubMed: 25253740) »

See all 13 Publications for this product

Customer reviews and Q&As

Thank you for contacting us. The two isoforms of macroH2A.1 vary in amino acids 198-229. The immunogen for this antibody does not overlap with this region and thus the antibody should react with both isoforms.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
Yes - TBST
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Username

Dr. Teo Manestar-Blazic

Verified customer

Submitted Sep 13 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Mouse Cell (Embryonic Stem Cells)
Specification
Embryonic Stem Cells
Preparation
Cell harvesting/tissue preparation method: feeder free single cell suspension via trypsination and gelatine culture for 1h
Sample buffer: PBS 0.3% Saponin
Fixation
Formaldehyde
Permeabilization
Yes - Saponin
Gating Strategy
life ES cell gate, excluding debris and MEFs
Username

Ms. Nadine Obier

Verified customer

Submitted Apr 28 2008

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

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