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.