Is it possible to blot the cell lysate with protein first then the antibody in western blot?

05 May 2018 1 5K Report

Hello there, I'm doing a western blot of my cells looking for binding of cd86 antibody. I am looking if my cells can bind the cd86 protein and then with the cd86 antibody. So is it possible to make the cells exposed to the cd86 protein first then check for anti-86 binding ?

Thanks

Bager

Santosh Jadhav

HI,

There are lot of discrepancies associated with it, mainly background signal. I would rather recommend far-western. Of course, you must isolate membrane proteins and also use cytosolic proteins as control (along with other controls). Check this publication

Article Detecting protein–protein interactions by Far Western blotting

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