negative control for IP

Amanda Lee leeam at umdnj.edu
Tue Jun 13 08:57:52 EST 2006


While doing immunoprecipitations with either Protein A Sepharose beads 
from GE or with the Upstate Catch and Release System my negative 
control IgG is pulling down many different proteins.  The bands that I 
get are equally as strong as in the lanes with my primary antibody.  I 
have tried blocking the beads in 5% BSA as well as preclearing the 
lysate before adding the antibodies but nothing has helped.

Does anyone know why I am getting so much non-specific binding in my 
IgG IPs?



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