Inclusion Bodies

Roger Murphy murphy_r at licre.ludwig.edu.au
Sun Aug 1 22:16:23 EST 1999

In article <37A1A4F8.29B8 at uctgsh1.uct.ac.za>, lester at uctgsh1.uct.ac.za wrote:
>I am currently trying to get proteins out of inclusion bodies in E.coli
>after an overnight culture growth and am really battling. Anyone out
>there with advice to get prots out of inclusion bodies without
>destroying/denaturing the proteins and rendering them inactive?
>Looking for help
>Dr L.Davids, Cape Town, South Africa
If your protein is in an inclusion body it's already likely to be denatured as 
it's not in a soluble form. 

The only way to get it out is to solubilise appropriately (we use 6M Gd.HCl, 
Tris buffer, pH 7, 10mM 2-ME) and then slowly remove the denaturing 
extractants by dialysis.

Fingers crossed that the protein then refolds as you want it!



Roger Murphy, Ph.D.
Biological Production Facility
Ludwig Institute for Cancer Research
Austin & Repatriation Medical Centre
Studley Road,
Heidelberg,  Vic. 3084

Tel  61-3-94965463
Fax  61-3-94965436
Email Roger.Murphy at Ludwig.edu.au

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