His tag and second purification step

Rover rover at livenet.net
Mon Apr 21 11:21:27 EST 1997


On Thu, 17 Apr 1997 16:34:42 +0100, "Koen A.L. De Smet"
<k.desmet at ic.ac.uk> wrote:

>I have a number of proteins with an N-terminal His-tag, expressed at low 
>level. Affinity purification on Ni++ enriches them, but it is still only 
>10% or so of the total protein. Also, I am convinced that I use enough 

Hello,
	What method are you using to purify, native (increasing
imidazole conc)?, denaturing (urea ,or guanidine)?. We have a few
proteins that form inclusion bodies so when using a none denaturing
(native) isolation procedure our yield is about 10% of total expressed
protein. Under denaturing conditions we genarally arrive at 90-100%
with no 2nd purification necessary.

 
Rover
Eastern Virginia Medical School



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