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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K79ac ChIP assays were performed on sheared chromatin from HeLa cells using 5 μg of the Diagenode monoclonal antibody against H3K79ac (cat. No. C15210006). QPCR was performed with primers for the coding region of the RPL30 gene, used as positive control and for the inactive MYOD1 gene and the Sata 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).
Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K79ac ChIP was performed on sheared chromatin from 1 million HeLa cells using 5 μg of the Diagenode antibody against H3K79ac (Cat. No. C15210006) 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. Western blot analysis using the Diagenode monoclonal antibody directed against H3K79ac 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 H3K79ac (cat. No. C15210006) diluted 1:1,000 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 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K79ac HeLa cells treated with butyrate were stained with the Diagenode antibody against H3K79ac No. C15210006 (red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).
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.
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Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
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