Anti-Histone H2B (phospho S32) antibody (ab10476)

This fast track antibody is not yet fully characterized. It is subject to these terms and conditions.

Overview

  • Product name
    Anti-Histone H2B (phospho S32) antibody
    See all Histone H2B primary antibodies
  • Description
    Rabbit polyclonal to Histone H2B (phospho S32)

    This product is a fast track antibody. It has been affinity purified and shows high titre values against the immunizing peptide by ELISA. Read the terms of use »

  • Host species
    Rabbit
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H2B, phosphorylated at S32.

    Read Abcam's proprietary immunogen policy (Peptide available as ab18504.)

  • General notes


    Serine 32 of Histone H2B was highlighted as being highly conserved in Cheung et al, indicating that it might be a good candidate phosphorylation site.

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
  • Primary antibody notes
    Serine 32 of Histone H2B was highlighted as being highly conserved in Cheung et al, indicating that it might be a good candidate phosphorylation site.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Fast track antibodies constitute a diverse group of products that have been released to accelerate your research, but are not yet fully characterized. They have all been affinity purified and show high titre values against the immunizing peptide (by ELISA). Fast track terms of use

Application Abreviews Notes
ICC/IF 1/200.
ELISA Use at an assay dependent concentration. : This antibody gave a positive result in ELISA against the immunizing peptide (ab18504).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/500. Detects a band of approximately 15 kDa (predicted molecular weight: 14 kDa).

Target

  • Relevance
    Core component of nucleosome. 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. Subunit structure The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Post-translational modification Monoubiquitination at Lys-35 (H2BK34Ub) by the MSL1/MSL2 dimer is required for histone H3 'Lys-4' (H3K4me) and 'Lys-79' (H3K79me) methylation and transcription activation at specific gene loci, such as HOXA9 and MEIS1 loci. Similarly, monoubiquitination at Lys-121 (H2BK120Ub) by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II. H2BK120Ub also acts as a regulator of mRNA splicing: deubiquitination by USP49 is required for efficient cotranscriptional splicing of a large set of exons. Phosphorylation at Ser-37 (H2BS36ph) by AMPK in response to stress promotes transcription. Phosphorylated on Ser-15 (H2BS14ph) by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination. GlcNAcylation at Ser-113 promotes monoubiquitination of Lys-121. It fluctuates in response to extracellular glucose, and associates with transcribed genes. Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes.
  • Cellular localization
    Nuclear
  • Database links
  • Alternative names
    • H2B GL105 antibody
    • H2B histone family member O antibody
    • H2B histone family member S antibody
    • H2B.1 antibody
    • H2B.1 B antibody
    • H2B.b antibody
    • H2B.c antibody
    • H2B.d antibody
    • H2B.e antibody
    • H2B.f antibody
    • H2B.j antibody
    • H2B.q antibody
    • H2B/b antibody
    • H2B/c antibody
    • H2B/d antibody
    • H2B/e antibody
    • H2B/f antibody
    • H2B/j antibody
    • H2B/o antibody
    • H2B/q antibody
    • H2BFB antibody
    • H2BFC antibody
    • H2BFD antibody
    • H2BFE antibody
    • H2BFF antibody
    • H2BFJ antibody
    • H2BFO antibody
    • H2BFQ antibody
    • H2BFS antibody
    • HIRIP2 antibody
    • HIST1H2BB antibody
    • HIST1H2BD antibody
    • HIST1H2BH antibody
    • HIST1H2BL antibody
    • HIST1H2BM antibody
    • HIST1H2BN antibody
    • HIST2H2BE antibody
    • Histone H2B antibody
    • Histone H2B type 1 B antibody
    • Histone H2B type 1 D antibody
    • Histone H2B type 1 H antibody
    • Histone H2B type 1 L antibody
    • Histone H2B type 1 M antibody
    • Histone H2B type 1 N antibody
    • Histone H2B type 2 E antibody
    • histone protein antibody
    see all

Images

This Fast-Track antibody is not yet fully characterised. These images represent inconclusive preliminary data.

  • All lanes : Anti-Histone H2B (phospho S32) antibody (ab10476) at 1/500 dilution

    Lane 1 : Hela control
    Lane 2 : Gamma irraditaed hela 1h
    Lane 3 : Gamma irraditaed hela 2h
    Lane 4 : Hela control with Human Histone H2B (phospho S32) peptide (ab18504) at 1 µg/ml
    Lane 5 : Gamma irraditaed hela 1h with Human Histone H2B (phospho S32) peptide (ab18504) at 1 µg/ml
    Lane 6 : Gamma irraditaed hela 2h with Human Histone H2B (phospho S32) peptide (ab18504) at 1 µg/ml
    Lane 7 : Hela control with Human Histone H2B peptide (ab18507) at 1 µg/ml
    Lane 8 : Gamma irraditaed hela 1h with Human Histone H2B peptide (ab18507) at 1 µg/ml
    Lane 9 : Gamma irraditaed hela 2h with Human Histone H2B peptide (ab18507) at 1 µg/ml

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 14 kDa
    Observed band size: 15 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Western blot using ab10476 at 1/500 on HeLa lysates taken at various time points after gamma irradiation (ab13823).

  • ICC/IF image of ab10476 stained HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab10476, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HepG2, Hek293 and MCF7 cells fixed in 4% PFA at 1ug/ml and Hela, Hek293, HepG2 and MCF7 cells fixed in 100% methanol at 1ug/ml. However, this Fast-Track antibody is not yet fully characterised. This image represents inconclusive preliminary data.
  • IHC image of ab10476 staining in Human Breast adenocarcinoma formalin fixed paraffin embedded tissue 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 ab10476, 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • ab10476 (1/200) staining Histone H2B phospho S32 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). for further experimental details please refer to Abreview.

    See Abreview

References

This product has been referenced in:
  • Li J  et al. Nuclear PKC-? facilitates rapid transcriptional responses in human memory CD4+ T cells through p65 and H2B phosphorylation. J Cell Sci 129:2448-61 (2016). WB, ICC/IF ; Human . Read more (PubMed: 27149922) »
  • Lau AT  et al. Phosphorylation of histone H2B serine 32 is linked to cell transformation. J Biol Chem : (2011). WB, ICC/IF ; Human . Read more (PubMed: 21646345) »
See all 3 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100

Dr. Kirk Mcmanus

Verified customer

Submitted Sep 08 2011

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

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