GST-fusion elution problem

MAEDA, Tatsuya, Ph.D. maeda at
Thu Sep 27 06:48:53 EST 2001

Dear All,

We have expressed a GST-fusion protein in bacteria.  Fortunately, the
protein was present in the supernatant following centrifugation of the
lysate.  Using glutathione beads, the protein was successfully recovered
from the supernatant.  And here comes a problem.  The protein resists to be
eluted from the beads.  We have tried several elution solutions: 10mM
glutathione 50mM Tris-Cl pH8.0, higher glutathione conc.(up to 50mM),
higher pH(pH9.6), detergent(0.2% Triton X-100), and higher salt conc.(up to
500mM NaCl), but all in vain.  Has anyone come across the same situation
and does anyone know any remedies?

Many thanks in advance.



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