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It is recommended that 30 mg of liver tissue is used for each ChIP/antibody. However, this amount may vary for other tissues. The exact amount of tissue depends upon protein abundance, antibody affinity and the efficiency of cross-linking. The protocol was optimized using 5-15 µg chromatin for each ChIP assay. The exact chromatin concentration should be determined for each tissue type before starting the X-ChIP assay. Our cross-linking chromatin immunoprecipitation (X-ChIP) protocol should be used after the chromatin preparation detailed below. Protease inhibitors should be included in all solutions used, including PBS PMSF 10 µl/ml, aprotinin 1 µl/ml and leupeptin 1 µl/ml).
This section was adapted from protocols kindly provided by Henriette O'Green, Luis G. Acevedo and Peggy J Farnham.
Please continue with the X-ChIP protocol from the sonication step
Adapted from protocols kindly provided by Henriette O’Geen, Luis G. Acevedo and Peggy J. Farnham.