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Problem with nonspecific antibody binding

Dr E. Buxbaum EB15 at le.ac.uk
Thu Feb 27 06:12:24 EST 1997

divernlb at aol.com (DiverNLB) wrote:
 The problem I am having is nonspecific binding of the
>secondary antibody (conjugated to fluorescein), leading to strong
>false-positive signals. I have tried blocking with bovine serum albumin,
>fetal bovine serum, and horse serum, with little or no effect. 

You could try to include fish skin gelatine (Sigma) into your solutions, 
this sometimes helps. Be warned though, this is a realy messy (though not 
dangerous) material. 

Are you sure that it is realy the secondary, rather than the primary 
antibody that is causing the problem? In this case you may want to change 
the source of the antibody (different company, different species). You 
may even label the primary antibody directly, if you have enough of it. 

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