Chromatin Immunoprecipitation Assay

After cross-linking the cells using 4% formaldehyde, the cells were disrupted by exposure to ultrasonic waves to obtain 200–1000 bp DNA–protein complex. Myelocytomatosis oncogene antibody was added to harvest DNA–MYC complex targeted by MYC antibody. DNA–MYC complex was incubated with Protein A Agarose/SaLmon Sperm DNA was supplemented to bind to MYC antibody-MYC-ENC1 promoter complex. These complexes were precipitated and rinsed to remove non-specific binding. The enriched MYC-ENC1 complexes were isolated and de-crosslinked. The enriched ENC1 promoter fragments were subjected to RT-qPCR analysis after purification.