H3K56ac polyclonal antibody

Lot	A2068P
Concentration	0.6 µg/µl
Species reactivity	Human
Type	Polyclonal
Purity	Affinity purified
Host	Rabbit
Precautions	This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Applications	Suggested dilution	References
ChIP/ChIP-seq ^*	5 μg/ChIP	Fig 1, 2
ELISA	1:500	Fig 3
Dot Blotting	1:20,000	Fig 4

H3K56ac Antibody ChIP Grade

Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K56ac (Cat. No. C15410213) 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 promoter and for the EIF4A2 promoter, 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).

H3K56ac Antibody ChIP-seq Grade

H3K56ac Antibody for ChIP-seq

H3K56ac Antibody for ChIP-seq assay

H3K56ac Antibody validated in ChIP-seq

Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac
ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 5 μg of the Diagenode antibody against H3K56ac (cat. No. C15410213) 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 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.

H3K56ac 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 H3K56ac (Cat. No. 15410213) 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 2), the titer of the antibody was estimated to be 1:15,300.

H3K56ac Antibody validated in Dot Blot

Figure 4. Cross reactivity tests using the Diagenode antibody directed against H3K56ac
To test the cross reactivity of the Diagenode antibody against H3K56ac (Cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. 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. Figure 3 shows a high specificity of the antibody for the modification of interest.

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.

実験手法

ELISA
Enzyme-linked immunosorbent assay. 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)
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ChIP-qPCR (ab)
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資料

Datasheet H3K56ac C15410213 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...	Download
Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...	Download