Dear Skivesh,
My favourite media is TPB - (paper is: Moore et al Protein expression
and purification (1993) 4: 160-163). I've had luck with this and
BL21 (DE3) Gold cells. I'm also a fan of cold shocking cells - after
initial growth at 37 deg C, put flasks in an ice water bath for 10
min, then add IPTG and tranfer to 16-25 deg C for expression. This
combination rescued a protein that behaved like ball bearings in all
other conditions. It always seems worth trying an array of strains
vs an array of media vs cold shock/heat shock/chaperones, and a bit
of tweaking of [IPTG]
Good luck,
Lou
At 11:06 02/04/2008, you wrote:
Dear all,
Sorry for the off-topic question...
What can be done to avoid a protein going inside inclusion body.The
gene is cloned in pET30a with C-ter his tag and expressed in
BL21-DE3 from 37 to 18C for 3-4 hr with .5mM of IPTG,it is going to
inclusion body.All suggestions are welcome.
Thanx in advance.
Shivesh
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Centre for Biomolecular Sciences
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