Inclusion body

Michael Witty mw132 at mole.bio.cam.ac.uk
Thu Oct 5 08:54:14 EST 2000


Scott, one of my collegues recommended sonication to resuspend.  I thought
it was too violent for a protein.  What do you think?  Mike.

On Wed, 4 Oct 2000, Scott Coutts wrote:

> 
> You could try using 6M guanidinium hydrochoride. I use a buffer with the
> following components:
> 
> 	6M Guanidinium.HCl
> 	20mM Tris.HCl
> 	100mM NaCl
> 	5mM DTT
> 
> Clean up your pellet (I assume you have the inclusion bodies ready for
> solubilisation), then very thoroughly resuspend it in the above buffer,
> making sure there are no 'lumps' (vortex, at least). Leave it overnight
> at four degrees celcius.
> 
> Hope that helps,
> 
> 
> Scott J. Coutts
> 
> Department of Microbiology
> Monash University
> Australia
> 
> 
> 
> 
> 
> Aki Hoji wrote:
> > 
> > Dear,
> > 
> > I tried to dissolve the purified inclusion body in the urea buffer (8M Urea,
> > 25mM MES, 1mM DTT 0,5mM EDTA) at room temp. overnight on the rotor but it
> > did not.  I'd appreciate any suggestions or comments.  Thanks in advance.
> > 
> > regards,
> > 
> > Aki Hoji
> > University of Pittsburgh
> > Department of Infectious Diseases and Microbiology
> > Rm522 Parran Hall, GSPH-IDM
> > 130 Desoto Street, Pittsburgh PA 15261
> > tel: (412) 624-3072/0776
> > fax: (412) 624-4953
> 
> 






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