GST fusion proteins HELP
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.
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