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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
A. B. C. D.
Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) 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 50 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 human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).
Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.
Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).
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A MORC-driven transcriptional switch controls Toxoplasma developmental trajectories and sexual commitment. Farhat DC, Swale C, Dard C, Cannella D, Ortet P, Barakat M, Sindikubwabo F, Belmudes L, De Bock PJ, Couté Y, Bougdour A, Hakimi MA Toxoplasma gondii has a complex life cycle that is typified by asexual development that takes place in vertebrates, and sexual reproduction, which occurs exclusively in felids and is therefore less studied. The developmental transitions rely on changes in the patterns of gene expression, and recent studies have assi...