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|-It is highly critical that the chromatin is of appropriate size and concentration.|
|-For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP.
-It is important to keep the tissue cold to avoid protein degradation.
-Use fresh formaldehyde that is not past the manufacturer's expiration date.
|-Once in solution, store 1M DTT at -20°C.|
|ChIP-exo was performed as previously described6,20 with chromatin extracted from 50 million cells, ProteinG MagSepharose resin (GE Healthcare), and 5 ug of antibody. Two biological replicates were prepared for each ChIP target. To enable future comparisons to drug treatment experiments, cells were treated with vehicle (final 0.03% DMSO (dimethyl sulfoxide)) for 4 h prior to harvest.|