Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9359(2)] to mH2A1
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-mH2A1 antibody [EPR9359(2)]
See all mH2A1 primary antibodies
DescriptionRabbit monoclonal [EPR9359(2)] to mH2A1
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human mH2A1 aa 200-300. The exact sequence is proprietary.
Database link: O75367
- 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.
Previously labelled as macroH2A.1.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
- Anti-mH2A1 antibody [EPR9359(2)] (Alexa Fluor® 488) (ab208563)
- Anti-mH2A1 antibody [EPR9359(2)] (Alexa Fluor® 647) (ab208879)
- Anti-mH2A1 antibody [EPR9359(2)] (HRP) (ab209320)
- Anti-mH2A1 antibody [EPR9359(2)] (Alexa Fluor® 555) (ab211851)
- Anti-mH2A1 antibody [EPR9359(2)] (Alexa Fluor® 594) (ab217090)
- Anti-mH2A1 antibody [EPR9359(2)] - BSA and Azide free (ab232602)
KO cell lines
KO cell lysates
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.
|WB||1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 1 µg/ml.
This antibody gives positive signal in both 4%PFA and 100% MeOH-fixed cells.
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 - 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).
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.
All lanes : Anti-mH2A1 antibody [EPR9359(2)] (ab183041) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : H2AFY knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Lanes 1- 2: Merged signal (red and green). Green - ab183041 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab183041 was shown to react with mH2A1 in wild-type HEK-293T cells in western blot. The band observed in knockout cell line ab266241 (knockout cell lysate ab257463) lane below 40kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and H2AFY knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab183041 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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.
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 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).
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 kidney tissue labeling mH2A1 with ab183041 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
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.
ab183041 has been referenced in 5 publications.
- 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). 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 . PubMed: 29800566
- Schueder F et al. Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT. Nat Commun 8:2090 (2017). ICC, STED micro ; Human . PubMed: 29233999
- Brighton PJ et al. Clearance of senescent decidual cells by uterine natural killer cells in cycling human endometrium. Elife 6:N/A (2017). PubMed: 29227245
- Fu Y et al. MacroH2A1 associates with nuclear lamina and maintains chromatin architecture in mouse liver cells. Sci Rep 5:17186 (2015). WB, ICC/IF . PubMed: 26603343