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Gal4 BD Antibody Problem

yuan zhou zhou at botany.utoronto.ca
Thu May 10 13:38:44 EST 2001


After yeast two-hybrid, we identified several cDNAs that encode for
proteins interacting with the bait.  Then we moved on to Western, hoping
to see those proteins bind the bait in vitro.  First, we separated
samples that contains AD-fused proteins, and then used the BD-bait as
probe.  However, we could not detect any signal using BD mAb from
Clontech.  The signal was missing even when the BD mAb was separated on
gel, and then detected with  the anti-mouse IgG.  Has anyone encountered
the same/similar problem?  Is the BD mAb not working?

Best regards,

Zhou




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