Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade – BSA and Azide free (ab237964)

Overview

  • Product name

    Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade – BSA and Azide free
    See all Histone H2B primary antibodies
  • Description

    Mouse monoclonal [mAbcam 52484] to Histone H2B - ChIP Grade – BSA and Azide free
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human Histone H2B aa 100 to the C-terminus (C terminal).

  • Positive control

    • ChIP: HeLa cells; WB: HeLa, NIH 3T3 and PC12 cells, zebrafish brain, heart, liver and skeletal muscle homogenates; ICC/IF: HeLa cells; IHC-P: Human breast tissue; IP: HeLa whole cell extract.
  • General notes

    Ab237964 is a PBS only version of ab52484.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at +4°C. Do Not Freeze.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    mAbcam 52484
  • Myeloma

    Sp2/0
  • Isotype

    IgG1

Applications

Our Abpromise guarantee covers the use of ab237964 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
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 14 kDa).
IHC-P Use a concentration of 1 µg/ml.
Flow Cyt Use 1µg for 106 cells.
ChIP Use 5 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

  • 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, 5µg of  ab52484 (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.  

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • All lanes : Anti-KDM4C / GASC1 / JMJD2C antibody (ab52584) at 5 µg/ml

    Lane 1 : Histone H1 recombinant protein
    Lane 2 : Histone H2A recombinant protein
    Lane 3 : Histone H2B recombinant protein
    Lane 4 : Histone H3 recombinant protein
    Lane 5 : Histone H4 recombinant protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Rabbit polyclonal to Mouse IgG - H&L (HRP) at 1/3000 dilution

    Performed under reducing conditions.

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



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • ICC/IF image of ab52484 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab52484, 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-mouse 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • IHC image of Histone H2B staining in human breast carcinoma FFPE section, performed on a 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 ab52484, 1µg/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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • Histone H2B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Histone H2B and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52484.
    Secondary: Protein G-HRP at 1/500 dilution.
    Band: 17kDa; Histone H2B; non specific bands - 26kDa: We are unsure as to the identity of this extra band.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • All lanes : Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 14 kDa


    Exposure time: 1 minute


    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • Overlay histogram showing HeLa cells stained with ab52484 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52484, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

  • Lanes 2-6 : Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 5 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish heart homogenate at 10 µg
    Lane 4 : Zebrafish liver homogenate at 10 µg
    Lane 5 : Zebrafish skeletal muscle homogenate at 10 µg
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Lanes 2-6 : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 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: 1 minute


    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).

References

ab237964 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab237964.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up