Overview

  • Product name

    Anti-Histone H2B (acetyl K15) antibody - ChIP Grade, purified
    See all Histone H2B primary antibodies
  • Description

    Rabbit polyclonal to Histone H2B (acetyl K15) - ChIP Grade, purified
  • Host species

    Rabbit
  • Specificity

    Due to sequence homology we expect this antibody to detect K15 acetylation on multiple H2B variants.
  • Tested applications

    Suitable for: ICC/IF, Dot blot, CHIPseq, WB, ChIPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Histone H2B (acetyl K15) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary. (The region of Histone H2B containing the acetylated lysine 15 (H2BK15ac)).
    Database link: Q5QNW6

  • Positive control

    • Whole cell and histone extracts from Hela cells; chromatin from Hela cells and HeLaS3 cells; recombinant histone H2A, H2B, H3 and H4. HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab195652 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 1/500.
Dot blot 1/20000.
CHIPseq Use at an assay dependent concentration.

Use 2 µg of antibody per 1.5x106 cells.

 

WB 1/500. Predicted molecular weight: 14 kDa.
ChIP Use at an assay dependent concentration.

Use 0.5-5 µg of antibody per 1.5x106 cells.

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

  • CHIP analysis of Histone H2B type 1-C/E/F/G/I (acetyl K15) in HeLa cells using ab195652 at 2 μg/IP, and optimized PCR primer sets for qPCR. ChIP was performed with the kit on sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. This image shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

     

  • ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 2 µg of ab195652, as described for the CHIP application. The IP’d DNA was subsequently analysed on a sequencer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the Human genome using the BWA algorithm. This image shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (sections A and B) and in genomic regions of chromosome 7, surrounding the ACTB gene, and of chromosome 12, surrounding the GAPDH gene (sections C and D). The position of the amplicon used for ChIPqPCR is indicated by an arrow.

     

  • To test the cross reactivity of ab195652, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified Histone H2B type 1-C/E/F/G/I. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1/20,000. This image shows a high specificity of the antibody for the modification of interest.

  • All lanes : Anti-Histone H2B (acetyl K15) antibody - ChIP Grade, purified (ab195652) at 1/500 dilution

    Lane 1 : whole extract from Hela cell at 25 µg
    Lane 2 : histone extract from Hela cell at 15 µg
    Lane 3 : recombinant histone H2A at 1 µg
    Lane 4 : recombinant histone H2B at 1 µg
    Lane 5 : recombinant histone H3 at 1 µg
    Lane 6 : recombinant histone H4 at 1 µg

    Predicted band size: 14 kDa



    ab195652 was diluted in TBS-Tween containing 5% skimmed milk.

  • Immunofluorescent analysis of Histone H2B type 1-C/E/F/G/I (acetyl K15) in HeLa cells using ab195652 at a 1/500 dilution in blocking solution, followed by an anti-rabbit antibody conjugated to Alexa488 (left). The cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/ TX-100 containing 5% normal goat serum and 1% BSA. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

References

ab195652 has not yet been referenced specifically in any publications.

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