Also worth trying a lower IPTG level ~ 0.1mM and or different media,
e.g. TB, SOC or m9 instead of LB, it's all a bit random but sometimes
these things can make a difference. Adding 3% EtOH on induction
worked for me once, it's supposed to shock the cells and stimulate
chaperone production, I was surprised I must say. Funny old world......
Giles Robertson D.Phil.
[EMAIL PROTECTED]
Tel: 02076316813
School of Crystallography,
Birkbeck College,
University of London,
Malet Street,
London, WC1E 7HX.
On 2 Apr 2008, at 12:19, Brenda Patterson wrote:
Lower temperature, use chaperones (e.g. TAKARA set), refolding?
Quoting shivesh kumar <[EMAIL PROTECTED]>:
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
