Re: [ccp4bb] Inclusion body

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

Xtal community:

- CCP4BB

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Nature policy update regarding source code
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-03-26

Next message:
Subject: Re: Nature policy update regarding source code
From: Ian Tickle I {- dot -} Tickle {- at -} ASTEX-THERAPEUTICS {- dot -} COM
Date: 2007-03-26

Subject: Re: Inclusion body
From: Cynthia Kinsland clk10 {- at -} CORNELL {- dot -} EDU
Date: 2007-03-26

We have tried them and had them work for a few proteins that didn't
look hopeful other ways.

The chaperones express well and obviously by gel and, in fact, we had
one protein that was difficult to separate from the Cpn60.

We very carefully follow the instructions on induction that come with
the cells (they are a little odd) and do the expression at 10°C.

We have added the ArcticExpress into our standard list of things to
try for rescuing insoluble proteins. We just started testing it out
a few weeks ago and it hasn't saved all of problem proteins by any
means. I don't have the numbers right now, but I think we got
something like 3 of 10 that looked insoluble at 15°C in BL21.

Best of luck,

Cynthia

On Mar 24, 2007, at 9:51 AM, Hongmin Li wrote:

> Hi Weijun and all,
>
> Anyone tried the Stratagene's ArcticExpress™ Competent Cells for
> expression of proteins at low temperature (13 degree). The company
> claims that the cells encode chaperonins Cpn60 and Cpn10 and they
> can enhance the protein folding and solubility.
>
> Hongmin
>
> >Dear All,
>
> >I got enclusion body in many cases when I tried to express human
> >proteins in E coli. I would like some suggestions on how to go about
> >it. I would also like to try co-expression of GroEL/GroES or
> >DnaJ/DnaK, and would like to know where to get the plasmids.
>
> >Any help or comments would be appreciated.
> >Weijun
>

____________________
Cynthia Kinsland, Ph.D.
Cornell University
Protein Facility Director
607-255-8844

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Nature policy update regarding source code
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-03-26

Next message:
Subject: Re: Nature policy update regarding source code
From: Ian Tickle I {- dot -} Tickle {- at -} ASTEX-THERAPEUTICS {- dot -} COM
Date: 2007-03-26