GST protein purification from inclusion bodies... - will GST part still be intact? (Apr/10/2006 )
since i failed to get much of soluble protein after induction in E-coli, i want to try to purify it from the inclusion body. But no protocol mentions the answer to my question: after using urea and all the stuff in the prification and solubilization buffers to solubilize the inclusion body, will i stil be able after that to attach my protein to GST beads? i mean will not all those chamicals effect GST part of my protein? thank you very much
Yes, the urea will denature your protein, and the GST tag needs to be folded properly in order to work. His tags are OK though. If you remove the urea, the GST tag may refold properly, but it may not.
thank you, i will remove urea by dialysis...do you think it is enough for GST refolding?