GST fusion rotein - (Nov/28/2001 )
Hi, I tries to elute the GST-fusion rotein from the Glutathione beads. It does not work well for me, even though I used freshly made elution buffer. What is the alternative way to do that?
I'm using GST fusion protein as well and to elute the proteins from the Glutathion beads: Buffer: 10mM Tris/HCl pH 7.5, 50mM NaCl, 0.1mM EDTA, 1mM DTT, 5mM Glutathion. After adding the glutathion, readjust the pH to 8 with solid Tris. You can try to let the column sit in elution buffer 10 mins before collecting the fractions. Usually, I have a good rate of recuperation.