I have performed an EMSA recently and have observed nice binding to my probe. When I added antibody to try to identify the protein binding the probe, I saw a decrease in band intensity, however I do not see a supershift of the complex. I have tried adding different amounts of antibody, but I am still unable to see a supershift. This antibody has been used to EMSA before. How do I optimise the assay for the supershift? And, does the decrease in band intensity when I add antibody, really suggest that the antibody is binding the complex?
Thanks a lot!!!
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problem with gel supershift
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