pGEX expression problems

farleyp at HAL.HAHNEMANN.EDU farleyp at HAL.HAHNEMANN.EDU
Wed Jun 23 13:03:02 EST 1993


The doublet protein may be an E. coli protein (dnaK) that co-purifies with many proteins expressed in pGEX-2T (is your extra band about 70 kDa?) The dnaK
protein runs around 70kDa on SDS-PAGE.  This protein purifies with glutathione-
agarose and I also have it co-purify when I use an immunoaffinity column.  I
understand that many people have had problems cleaving their fusion protein
with thrombin.  Good luck --

Patrick Farley
farleyp at hal.hahnemann.edu



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