Inclusion body

Scott Coutts scott.coutts at med.monash.edu.au
Wed Oct 4 02:52:20 EST 2000


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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