[Histonet] RE: Negative Immunohistochemistry controls

From:"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 o=66 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.


==========================3D=========================3D=========================3D==
The contents of this message, together with any attachments, are intended only for the use of the person(s) to which they are addressed and may contain confidential and/or privileged information. Further, any medical information herein is confidential and protected by law. It is unlawful for unauthorized persons to use, review, copy, disclose, or disseminate confidential medical information. If you are not the intended recipient, immediately advise the sender and delete this message and any attachments. Any distribution, or copying of this message, or any attachment, is prohibited.
==========================3D=========================3D=========================3D==


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


<< Previous Message | Next Message >>