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GST protein batch purification

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Posted 18 July 2011 - 04:26 AM

Hi every body,

I have a funny problem regarding eluting my GST bind protein: the purification is a batch method by glotathion sepharos 4b beeds and after GST-resin binding there is thrombin cleavage to release the purified protein and then I spin down the resin beeds and take the supernatant which contains my protein. the problem is despite lots of washes (with thrombin buffer pH 7.5) there are still a lot of protein remaining in the resin beeds they are cuted off properly (as SDS-PAGE shows but kind of trapped between beeds. any suggestion?

Many thanks,

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