Inclusion Bodies

[Roger Murphy]

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!

Cheers,

Roger


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

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