Diagenode

H2BK12ac Antibody

Catalog Number
Format
Price
C15410212
50 μg
$380.00
  Bulk order
Other format



Polyclonal antibody raised in rabbit against the region of histone H2B containing the acetylated lysine 12 (H2BK12ac), using a KLH-conjugated synthetic peptide.

LotA2260P
Concentration0.82 µg/µl
Species reactivityHuman
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 0.5 - 1 μg per ChIP Fig 1, 2
ELISA 1:1,000 Fig 3
Dot Blotting 1:5,000 Fig 4
Western Blotting 1:1,000 Fig 5
Immunofluorescence 1:500 Fig 6

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 μg per IP.

  • Validation Data

    H2BK12ac Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H2BK12ac
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2BK12ac (Cat. No. C15410212) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using 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. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H2BK12ac Antibody ChIP-seq Grade

    H2BK12ac Antibody for ChIP-seq

    H2BK12ac Antibody for ChIP-seq assay

    H2BK12ac Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H2BK12ac
    ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 0.5 μg of the Diagenode antibody against H2BK12ac (Cat. No. C15410212) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. 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. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 7, surrounding the ACTB gene, and of chromosome 12, surrounding the GAPDH gene (fig 2C and D). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.

    H2BK12ac Antibody ELISA validation

    Figure 3. Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2BK12ac (Cat. No. C15410212) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:38,200.

    H2BK12ac Antibody validated in Dot Blot

    Figure 4. Cross reactivity tests using the Diagenode antibody directed against H2BK12ac
    To test the cross reactivity of the Diagenode antibody against H2BK12ac (Cat. No. C15410212), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H2B. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

    H2BK12ac Antibody validated in Western Blot

    Figure 5. Western blot analysis using the Diagenode antibody directed against H2BK12ac
    Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H2BK12ac (Cat. No. C15410212). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left.

    H2BK12ac Antibody validated in Immunoflurescence

    Figure 6. Immunofluorescence using the Diagenode antibody directed against H2BK12ac
    HeLa cells were stained with the Diagenode antibody against H2BK12ac (cat. C15410212) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2BK12ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • Target Description

    Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of histone H2B is associated with active genes.

  •  Applications
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    WB
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
    IF
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
  •  Documents
    Datasheet H2BK12ac C15410212 DATASHEET
    Datasheet description
    Download
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
  •  Safety sheets
    SDS C15410212 H2BK12ac Antibody US en Download
    SDS C15410212 H2BK12ac Antibody GB en Download
    SDS C15410212 H2BK12ac Antibody DE de Download
    SDS C15410212 H2BK12ac Antibody ES es Download
    SDS C15410212 H2BK12ac Antibody FR fr Download
    SDS C15410212 H2BK12ac Antibody BE fr Download
    SDS C15410212 H2BK12ac Antibody JP ja Download
    SDS C15410212 H2BK12ac Antibody BE nl Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H2BK12ac Antibody (Diagenode Cat# C15410212 Lot# A2260P). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Identification of Novel Molecular Markers of Human Th17 Cells.
    Sałkowska A, Karaś K, Karwaciak I, Walczak-Drzewiecka A, Krawczyk M, Sobalska-Kwapis M, Dastych J, Ratajewski M
    Th17 cells are important players in host defense against pathogens such as , , and . Th17 cell-mediated inflammation, under certain conditions in which balance in the immune system is disrupted, is the underlying pathogenic mechanism of certain autoimmune disorders, e.g., rheumatoid arthritis, Graves' disease, multi...

    Myc Regulates Chromatin Decompaction and Nuclear Architecture during B Cell Activation
    Kieffer-Kwon K.R. et al.
    50 years ago, Vincent Allfrey and colleagues discovered that lymphocyte activation triggers massive acetylation of chromatin. However, the molecular mechanisms driving epigenetic accessibility are still unknown. We here show that stimulated lymphocytes decondense chromatin by three differentially regulated steps. Fi...

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