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IHC Protein Competition, Protein Background

IHC Protein Competition Background Blocking

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#1 Laura Robin

Laura Robin

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Posted 12 October 2016 - 12:25 PM

Hi there,

 

I am attempting to perform a protein competition with a commercial antibody. The signal from the antibody seems to have been reduced, but we cannot say definitively as there is intense background from the protein on the tissue. 

 

I was hoping someone could recommend me the best course of action in order to eliminate this background. I have tried BSA, but this eliminates staining from my antibody.

 

Thanks for your help!



#2 bob1

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Posted 13 October 2016 - 01:14 PM

There are a range of different buffers you can use for antibody binding solutions (other than tris and phosphate based buffers), and also a range of different blocking reagents you might try (casein, gelatin (fish, pig, cow), normal antisera, albumin etc.). 

 

Some details on your protocol might help.







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