12CA5 MAb immunoprecipitation

Jean-Yves Masson massonjy at rsvs.ulaval.ca
Wed Aug 30 18:49:13 EST 1995

Dear Netters,

        We are trying to immunoprecipitate (from total yeast lysate) an 
epitope tagged protein. The epitope is the HA tag YPYDVPDYA with the MAb 
12CA5. We're having high background problems coming from non-specific 
binding to protein A-sepharose beads. The antibody worked great for 
western blot detection, but not at all for immunoprecipitation. My buffer 
contains 50 mM Tris pH 7.5, 150 mM Nacl, 2 mM MgCl2, 1 mM EDTA, 0.5 % 
NP-40 and 1 mM PMSF. 

	Does anyone have a protocol for immunoprecipitation with the 
12CA5 Ab ? Also, any suggestions, references or hints would be greatly 

Thanks in advance,

J-Y Masson
CHUL research center
Quebec, CANADA

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