raising ANTIBODIES with fusion proteins??

T. S. Pillay tpillay at ucsd.edu
Sat Sep 10 16:40:43 EST 1994


Subject: raising ANTIBODIES with fusion proteins??
From: Riccardo Fodde, fodde at ruly46
Date: Tue, 6 Sep 1994 17:49:12
In article <fodde.13.0011D278 at ruly46> Riccardo Fodde, fodde at ruly46 writes:
>We have made several his-tagged fusionproteins which we used to immunize 
>rabbits. etc.

The problem may lie with the antirabbit antibody you are using for
detection.  A lot of the ones available commercially react with things
that they're not supposed to react with- eg.  sometimes the molecular
weight markers are detected.  Have you excluded this as a problem by
incubating blots with the 2nd antirabbit antibody alone to see which
bands come up.  Also, you might want to couple your antibody directly to
peroxidase or whatever enzyme you're using.



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