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Customer kindly called to discuss whether they might use a purificed sample in lieu of nuclear sample. Specifically they wish to perform an IP with their sample elute the protein of interest using a FLAG tag and then running the Histone Acetyltransferase Activity Assay. Although they are aware this kit has not been tested in these conditions, they wish to know if this sample type might work and if by using this sample type they might have to adjust other parts of the protocol.
Asked on May 21 2012
I have had the chance to inquire to the lab about the use of a purified IP sample. Theywould suggest using the initial sample without any IP for the HAT assay. If the IP is just for concentrating the sample, they recommend use of spin columns, not IP.
Theoretically though, the IP samples should also work they do not have any experience using this method and though modifications to the protocol may have to be performed to optimize the reaction, these would have to be determine experimentally in the lab.
Answered on May 21 2012