immunoaffinity purification

yqhuang at iastate.edu yqhuang at iastate.edu
Wed May 4 14:49:29 EST 1994


Dear netters, 
I have been trying to purify a cell surface glycoprotein by immunoaffinty
chromatography.I tried protein G-sepharose for both purification and
covalent coupling  of the monoconal antibody.That worked just fine.I
solubilized the membrane proteins in detergent,without first purifying the
membrane.The problems were:1)elution:I tried high pH,low pH,high salt(4 M
MgCl2),NaSCN,urea,and even guanidine.HCl,no one satisfied me for recovery. 
2)purity.I used  proteinG-sepharose precolumn,but still could not get rid of
the contaminants.Lentil lectin column did not help either.  

Thank you for your advice in advance.

Yueqiao Huang
3178 Molecular Biology Bldg
Iowa State University
Ames,IA 50011
fax:515-294-0345
email:yqhaung at iastate.edu
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