GST fusion proteins HELP

sujoy ghosh sujoy_ghosh at glaxo.com
Tue Oct 22 14:18:37 EST 1996


In article <324AA269.D71 at warren.med.harvard.edu>,
berez at WARREN.MED.HARVARD.EDU wrote:

> I am trying to express a leech homeobox protein in bacteria to make an 
> antibody.  The protein is insoluble.  I followed the Frangioni and 
> Neel paper for solubilizing using sarkosyl.  I get a good amount of 
> the protein into the supernatant, then I add Triton to restore binding 
> to glutathione sepharose but I don't get a good yeild.  I get back the 
> tiniest fraction of what is in the supernatant.  I also seem to get 
> poor elution from the beads.  Any ideas?
> 
> Ashley Bruce


What are your growth conditions? Assuming you are growing your cultures at
37 degrees, try growing them at 20 degrees for 1-3 hours. It might help.

Sujoy



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