protein solubility - (Oct/20/2008 )
I'm going to purify a protein using Ni column and as I have read the optimum pH for this technique is about 8. unfortunately the pI of the protein which is the focus of my interest is 8 too which makes it insoluble at this pH. I wonder whether there is a way to get around this problem?
P.S. my protein is 145 kD
-hppkm.y-
QUOTE (hppkm.y @ Oct 20 2008, 05:43 PM)
I'm going to purify a protein using Ni column and as I have read the optimum pH for this technique is about 8. unfortunately the pI of the protein which is the focus of my interest is 8 too which makes it insoluble at this pH. I wonder whether there is a way to get around this problem?
P.S. my protein is 145 kD
P.S. my protein is 145 kD
It doesnt have to be exactly pH8.. I have used pH 6-8 where pH6 worked better with fewer contaminants than pH8... just try lower pH and see what happens.
-scFv-
HI
U don't have to bind protein at pH=8, u should look at the Ni column handbook the range of optimum working (binding protein).
it should also bind at pH=7 or less.
bye
-amtash-