Dear all, In continuation with my earlier mail on problem in cation exchange chromato, i was suggested to go for hydrobic interaction chromato [HIC] as an additional step. my protein is indeed a hydrophobic protein. we have sepharose 6 fast flow low sub resin and my protein has a pi of 5.8 -6.3. one of the patent papres relating to the same moleculte it was mentioned HIC in suitable buffer containing 0.5 M NH4SO4 pH of 5.0 and elution in same buffer without NH4SO4 pH 5.0. They have mentioned addition of 2-20% ethanol in the elution buffer. is it O.K. to add ethanol in buffer.
Can anyone provide me a suitable buffer recipe to use at pH 5.0 for HIC. i performed the HIC but the resolution was not good and my sample was diluted 10 fold and elution was near end of elution gradient. Kindly comment thanks in anticipation
