Re: [ccp4bb] "inclusion body"

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Subject: Re: "inclusion body"
From: Mohinder Pal mp11 {- at -} SOTON {- dot -} AC {- dot -} UK
Date: 2008-04-02

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From: "Mark J {- dot -} van Raaij" VANRAAIJ {- at -} USC {- dot -} ES
Date: 2008-04-02

Subject: Re: "inclusion body"
From: Giles Robertson g {- dot -} robertson {- at -} MAIL {- dot -} CRYST {- dot -} BBK {- dot -} AC {- dot -} UK
Date: 2008-04-02

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.
g.robertson@mail.cryst.bbk.ac.uk
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 <2shivesh@GMAIL.COM>:
>
>> 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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CCP4bb <-- 2008 <-- April 2008 <-- 02 April 2008

Previous message:
Subject: Re: "inclusion body"
From: Mohinder Pal mp11 {- at -} SOTON {- dot -} AC {- dot -} UK
Date: 2008-04-02

Next message:
Subject: Co-expression plasmids
From: "Mark J {- dot -} van Raaij" VANRAAIJ {- at -} USC {- dot -} ES
Date: 2008-04-02