Western specicity cross contamination

Nameer B Kirma nkirma at emory.edu
Thu Feb 26 11:51:52 EST 1998

I have been trying to detect different mouse  proteins using specific
mouse monoclonal antibodies. Both primary and secondary antibodies were
made in mice.  Besides the specific bands I am getting a thick 45 kd band
in all the blots.  Blotting with  only the secondary antibody gives
almost the same 45 kd band, which suggests it's crossreactivity with the
mouse IgG.  Are there ways to eliminate the nonspecific (background) band?
I have tried using a lower antibody dilution but that did not work.

Thanks for your input.    

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