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About Gel supershift


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#1 lipeiqiang78

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Posted 01 May 2013 - 08:54 AM

[font=宋]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?
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[font=宋]Thanks in advance!!![/font]



#2 lipeiqiang78

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Posted 01 May 2013 - 09:12 AM

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!!!


Edited by lipeiqiang78, 01 May 2013 - 09:14 AM.





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