About Gel supershift - (May/01/2013 )

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, which others in my lab tell me is indicative of the antibody binding the protein:probe complex, 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?


Thanks in advance!!!

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, which others in my lab tell me is indicative of the antibody binding the protein:probe complex, 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 in advance!!!