Key features and details
- Mouse monoclonal [14G7] to mH2A1
- Suitable for: ChIP, WB, ELISA, Flow Cyt
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-mH2A1 antibody [14G7]
See all mH2A1 primary antibodies
DescriptionMouse monoclonal [14G7] to mH2A1
Tested applicationsSuitable for: ChIP, WB, ELISA, Flow Cytmore details
Species reactivityReacts with: Human
Recombinant full length protein corresponding to Human mH2A1.
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; MCF7
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Concentration information loading...
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.
|ChIP||Use at an assay dependent concentration.|
|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.
FunctionVariant 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.
Sequence similaritiesContains 1 histone H2A domain.
Contains 1 Macro domain.
modificationsMonoubiquitinated 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 localizationNucleus. Chromosome. Enriched in inactive X chromosome chromatin and in senescence-associated heterochromatin.
- Information by UniProt
- Core histone macro h2a.1 antibody
- Core histone macro-H2A.1 antibody
- H2A histone family member Y antibody
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 - ab91528 observed at 40 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab91528 was shown to specifically recognize mH2A1 in wild-type HAP1 cells. No band was observed when mH2A1 knockout cells were examined. Wild-type and mH2A1 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.
Chromatin was prepared from SK-OV-3 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab91528 (red), or 5 µg of mouse normal IgG1 ab18443 (gray) and 25 µl of Protein A/G Dyna beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are commercial primers from Paper PMID: 22589551
All lanes : Anti-mH2A1 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.
All lanes : 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
ab91528 has not yet been referenced specifically in any publications.