Recombinant

Recombinant Anti-Histone H2B antibody [IGX4228H] (ab213288)

Overview

  • Product name

    Anti-Histone H2B antibody [IGX4228H]
    See all Histone H2B primary antibodies
  • Description

    Human monoclonal [IGX4228H] to Histone H2B
  • Host species

    Human
  • Tested applications

    Suitable for: WB, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human Histone H2B.

  • Positive control

    • WB: CTH, HeLa whole cell, HeLa Nuclear, NIH3T3 whole cell, NIH3T3 Nuclear and Histone H2B Recombinant Protein lysates. ChIP: HeLa and NIH3T3.
  • General notes

    This product was made using synthetic libraries and phage display technology.

    This antibody is a recombinant antibody. 
    Human monoclonal antibody.

    For recommended secondary antibodies -

    Rabbit monoclonal Anti-Human IgG1 H&L (Alexa Fluor® 488) - ab200622
    Rabbit monoclonal Anti-Human IgG1 H&L (Alexa Fluor® 647) - ab200623
    Rabbit monoclonal Anti-Human IgG1 H&L (Biotin) - ab201485
    Rabbit Anti-Human IgG H&L (HRP)  - ab6759

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Monoclonal
  • Clone number

    IGX4228H
  • Isotype

    IgG1

Applications

Our Abpromise guarantee covers the use of ab213288 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 Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 14 kDa).
ChIP Use 2 µg for 25 µg of chromatin.

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

    • GL105 antibody
    • 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
    • H2BGL105 antibody
    • H2BQ antibody
    • HIRIP2 antibody
    • HIST1H2BB antibody
    • HIST1H2BD antibody
    • HIST1H2BH antibody
    • HIST1H2BL antibody
    • HIST1H2BM antibody
    • HIST1H2BN antibody
    • HIST2H2BE antibody
    • histone cluster 2 antibody
    • Histone H2B antibody
    • histone H2B GL105 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 H2B.q antibody
    • histone protein antibody
    see all

Images

  • All lanes : Anti-Histone H2B antibody [IGX4228H] (ab213288) at 1 µg/ml

    Lane 1 : CTH (Calf Thymus Histone) at 0.5 µg
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear Lysate at 10 µg
    Lane 6 : Histone H2B Recombinant Protein at 0.1 µg
    Lane 7 : Histone H3 Recombinant Protein at 0.1 µg

    Secondary
    All lanes : HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 14 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab213288 overnight at 4°C. Antibody binding was detected using an anti-human antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab213288 (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). Primers and probes are located in the first kb of the transcribed region.

  • Chromatin was prepared from NIH3T3 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab213288 (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 (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.

References

ab213288 has not yet been referenced specifically in any publications.

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