Recombinant
RabMAb

Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886)

Overview

  • Product name
    Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade
    See all Histone H2B primary antibodies
  • Description
    Rabbit monoclonal [EP857Y] to Histone H2B (acetyl K5) - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    There is cross-reactivity with H3K27Ac (Histone H3 acetylated on Lys 27).
  • Tested applications
    Suitable for: WB, IHC - Wholemount, CHIPseq, ChIP, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Caenorhabditis elegans
  • Immunogen

    Synthetic peptide within Human Histone H2B aa 1-100 (acetyl K5). The exact sequence is proprietary.

  • Positive control
    • TSA-treated A431 cell lysate, human urinary bladder carcinoma.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab40886 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
WB 1/50000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
IHC - Wholemount Use at an assay dependent concentration.
CHIPseq Use at an assay dependent concentration.
ChIP Use 6-25 µg for µg of chromatin.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • 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

    • Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 6µl of ab40886 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). 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 located in the first kb of the transcribed region.    

    • All lanes : Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886) at 1/50000 dilution

      Lane 1 : A431 cell lysate - untreated
      Lane 2 : A431 cell lysate - treated with TSA

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 14 kDa
      Observed band size: 14 kDa

    • ab40886 staining human bladder carcinoma for Histone H2B expression (1:250 dilution)
    • ab40886 (1:250) staining A431 cells by immunofluorescence.
    • ab40886 at a 1/600 dilution for ChIP analysis of mouse dorsal skin epidermis whole tissue lysate, incubated for 15 hours at 4°C with ChIP dilution buffer. Cross-linking (X-ChIP) using 1% formaldehyde for 10 minutes.
      Detection step: Semiquantitative PCR.
      Negative control: Rabbit IgG.
      Cells treated with active vitamin D3.

      See Abreview

    • IHC - Wholemount of Caenorhabditis elegans larvae labelling Histone H2B (acetyl K5) with ab40886. The sample was incubated with primary antibody (1/200 in PBS + 3% BSA + 0.1% Triton X-100) for 12 hours at 4°C. ab150077, a goat anti-rabbit Alexa 488 (1/1000), was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:
    • Brocks D  et al. DNMT and HDAC inhibitors induce cryptic transcription start sites encoded in long terminal repeats. Nat Genet 49:1052-1060 (2017). Read more (PubMed: 28604729) »
    • Gao J  et al. HDAC3 But not HDAC2 Mediates Visual Experience-Dependent Radial Glia Proliferation in the Developing Xenopus Tectum. Front Cell Neurosci 10:221 (2016). IHC-Fr ; Xenopus laevis . Read more (PubMed: 27729849) »
    See all 6 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (12.5%)
    Sample
    Fruit fly (Drosophila melanogaster) Tissue lysate - whole (Fruit fly larvae)
    Specification
    Fruit fly larvae
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Jan 08 2015

    Application
    IHC - Wholemount
    Sample
    Caenorhabditis elegans Embryo (C. elegans larvae)
    Specification
    C. elegans larvae

    Abcam user community

    Verified customer

    Submitted Dec 30 2013

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 23°C
    Antigen retrieval step
    None
    Sample
    Rat Tissue sections (Rat olfactory bulb frozen section)
    Specification
    Rat olfactory bulb frozen section
    Permeabilization
    Yes - 0.1% Triton X-100
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 30 2013

    Application
    Immunocytochemistry
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 23°C
    Sample
    Human Cultured Cells (Human embryonic stem cells)
    Specification
    Human embryonic stem cells
    Permeabilization
    Yes - 0.3% Triton X-100
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 27 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    ChIP
    Sample
    Mouse Tissue lysate - whole (dorsal skin epidermis)
    Specification
    dorsal skin epidermis
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% formaldehyde
    Detection step
    Semiquantitative PCR
    Negative control
    Rabbit IgG

    Abcam user community

    Verified customer

    Submitted Feb 15 2010

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

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