[Histonet] RE: Negative Immunohistochemistry controls

[Kellar, Eric C]

Cynthia,

Negative controls are used to check the presence of non-specific staining. An adequate (-) control consists of replacing the specific primary antibody by another antibody produced in the same species but with unrelated specificity. When using polyclonal antisera, a preimmune serum may be used. With monoclonal antibodies, normal mouse serum or non-immune mouse ascites is applied. They are used to evaluate the non-specific binding of that isotope to the cells. They are applied in the highest concentration that is used for the specific antibodies. In indirect or multi-step techniques the primary antibody may also be omitted to evaluate the non-specific binding of the other reagents.

Eric C. Kellar
Histology/Immunohistochemistry
Quest Diagnostics, Inc Miami







Good Morning everyone, I have a question about immuno
staining. I've been away from this type of staining
for a while and now I am doing them again on a regular
basis. Why do you run a negative control with each
run? Are you only suppose to put the normal serum on
negative control only? I've forgotten; would someone
please answer these questions for me. Thanks in
advance.

Cynthia Haynes H.T.


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