GST-GFP Purification Problem

Scott-Navigator moyerowl at blue.weeg.uiowa.edu
Sat Feb 13 09:45:06 EST 1999


We are trying to purify a fusion protein consisting of GST with GFP
fused to its C-terminus.  The protein seems to be expressed fairly well
and is soluble.  The supernatant is green due to the high concentration
of GFP.  When sepharose-GSH beads are added, the supernatant loses its
green color and now the beads are green.  So far, so good.  Now the bad
part.  We can't seem to elute the fusion protein even with very high GSH
concentrations.  Other GST fusion proteins have been purified by us
using this same protocol.  Any advice/suggestions/wild guesses would be
much appreciated.

Thanks,
Scott Moye-Rowley




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