Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K79me2
ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K79me2 (cat. No. CS-051-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 7 and 15 μl per ChIP experiment was analysed. Additionally, ChIP was performed with 2 and 15 μl of antibody after incubation with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).