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Hi all, I've recently switched over to using a GST system for purifying my proteins. Although I can get enough protein out of a few liters, I've always been finding that when I take some of the beads to run on a gel, I always get a decent amount of protein just stuck on there that won't elute off no matter what. I don't know what proportion is just sticking, but I would love to get better yields. I'm purifying using a HEPES based buffer at pH8.0 with 350mM NaCl in the buffer. Does anyone else have any ideas as to what I can add to my buffers to help reduce the amount of sticking? Thanks, Peter