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Figure 1. ChIP results obtained with the Diagenode antibody directed against MYH11 ChIP assays were performed using ME-1 cells, the Diagenode antibody against MYH11 (Cat. No. C15310254) and optimized primer pairs for qPCR. Sheared chromatin from 1.5 million cells and 5 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the genes indicated. Figure 1 shows the relative occupancy, calculated as the ratio + control/background for which the MYOG gene was used.
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against MYH11 ChIP was performed as described above. The IP’d DNA from 6 ChIP’s was pooled and subsequently analysed on an Illumina HiSeq 2000. 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 signal in 4 genomic regions surrounding the AXIN1, FUT7, BCL3 and RAD50 positive control genes.
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 human MYH11 (Cat. No. C15310254). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:1,900.
Figure 4. Western blot analysis using the Diagenode antibody directed against MYH11 Nuclear extracts of ME-1 cells were analysed by Western blot using the Diagenode antibodies against CBFb (Cat. No. C15310002, lane 1) and MYH11 (cat. No. C15310254, lane 2) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the CBFb and CBFb-MYH11 fusion proteins is indicated on the right.
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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