Anti-mH2A1 antibody (ab37264)
Key features and details
- Rabbit polyclonal to mH2A1
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-mH2A1 antibody
See all mH2A1 primary antibodies -
Description
Rabbit polyclonal to mH2A1 -
Host species
Rabbit -
Specificity
ab37264 recognises the three known isoforms of mH2A1 including mH2A1.2 (longest isoform) and the mH2A1.1 (shortest isoform).
We have had varying reports about the efficiency with which this antibody recognises mH2A1 in mouse cells and tissues. Please contact our Scientific Support team if you have any queries about this. -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human mH2A1 aa 150-250 conjugated to keyhole limpet haemocyanin.
(Peptide available asab37263) -
General notes
Previously labelled as macroH2A.1.
Properties
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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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanIHC-P HumanWB HumanAll applications RatChickenCowXenopus laevisApplication Abreviews Notes WB (1) Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).IHC-P (1) Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.ICC/IF (1) Use a concentration of 1 - 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).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
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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
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 localization
Nucleus. Chromosome. Enriched in inactive X chromosome chromatin and in senescence-associated heterochromatin. - Information by UniProt
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Database links
- Entrez Gene: 9555 Human
- Entrez Gene: 26914 Mouse
- Entrez Gene: 29384 Rat
- Omim: 610054 Human
- SwissProt: O75367 Human
- SwissProt: Q9QZQ8 Mouse
- SwissProt: Q02874 Rat
- Unigene: 420272 Human
see all -
Alternative names
- Core histone macro h2a.1 antibody
- Core histone macro-H2A.1 antibody
- H2A histone family member Y antibody
see all
Images
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Western blot - Anti-mH2A1 antibody (ab37264)
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 - ab37264 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab37264 was shown to specifically recognize MH2A1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when MH2A1 knockout cells were examined. Wild-type and MH2A1 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.
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Immunocytochemistry/ Immunofluorescence - Anti-mH2A1 antibody (ab37264)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mH2A1 antibody (ab37264)IHC image of mH2A1 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.
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Western blot - Anti-mH2A1 antibody (ab37264)All lanes : Anti-mH2A1 antibody (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
All lanes : 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
Protocols
Datasheets and documents
References (19)
ab37264 has been referenced in 19 publications.
- Haque N et al. ZFR coordinates crosstalk between RNA decay and transcription in innate immunity. Nat Commun 9:1145 (2018). PubMed: 29559679
- Rona G et al. PARP1-dependent recruitment of the FBXL10-RNF68-RNF2 ubiquitin ligase to sites of DNA damage controls H2A.Z loading. Elife 7:N/A (2018). PubMed: 29985131
- Sun X et al. Ki-67 Contributes to Normal Cell Cycle Progression and Inactive X Heterochromatin in p21 Checkpoint-Proficient Human Cells. Mol Cell Biol 37:N/A (2017). PubMed: 28630280
- Fontanals-Cirera B et al. Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene. Mol Cell 68:731-744.e9 (2017). PubMed: 29149598
- Leung JW et al. ZMYM3 regulates BRCA1 localization at damaged chromatin to promote DNA repair. Genes Dev 31:260-274 (2017). PubMed: 28242625
Images
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Western blot - Anti-mH2A1 antibody (ab37264)
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 - ab37264 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab37264 was shown to specifically recognize MH2A1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when MH2A1 knockout cells were examined. Wild-type and MH2A1 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.
-
Immunocytochemistry/ Immunofluorescence - Anti-mH2A1 antibody (ab37264)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mH2A1 antibody (ab37264)
IHC image of mH2A1 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.
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Western blot - Anti-mH2A1 antibody (ab37264)All lanes : Anti-mH2A1 antibody (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
All lanes : 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
