Immunohistochemistry: Technique Question

Auditorium you at somehost.somedomain
Tue Mar 26 15:07:44 EST 1996


I'm doing some immunohistochemistry on paraformaldehyde fixed specimens.  
I have a question on the reagent that the primary and secondary antibodies 
are made in.  These antibodies are from Peninsula Laboratories (they are 
not on the Web, I've done a Lycos search).  The package insert says to mix 
these antibodies in a PBS/Triton-X solution.  There is not a mention of 
including a non-specific protein (such as non-fat milk or normal goat 
serum or BSA, etc.).  I'm curious as to why most of the other techniques 
using a primary and secondary antibody to detect a protein require that 
both of the antibodies to be mixed up in a fairly high concentration of a 
non-specific protein.  I've followed Peninsula's directions and my 
specimens had very poor signal to noise ratio.  Could it be that I should 
have made these antibodies in a protein solution?  Email me back at 
tcc5g at virginia.edu.

Thanks

T. Chai




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