GST Protein purification problems

Graham Taylor graham.taylor at bigfoot.com
Mon Nov 19 12:37:30 EST 2001


I'm currently trying to purify a GST tagged protein (100kD) expressed
in insect cells via baculovirus.  Protein is produced and is soluble,
but... fails to bind to the GST column.  The cells were lysed in the
presence of 5mM EDTA and I performed the binding in the cold room. 
Protein was eluted using glutathione. A small 'smear' of protein of
increased (!) size was observed in the eluted protein.
Has anyone else tackled this problem.  One final thing, the his tagged
version of this protein also failed to bind to a Talon column.  I'm
getting tired of all this purification, can anyone help?

Thanks for reading this post
graham




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