[Histonet] negative immunohistochemistry control
Rene J Buesa
rjbuesa <@t> yahoo.com
Wed Mar 8 07:51:35 CST 2006
Cynthia:
The idea of the negative control is to try to determine if any reaction detected in the case is due to specific or unspecific binding.
You don't need to run a negative slide for each antibody you are going to test on the case, you only need a negative slide per case, no matter how many antibodies you run. The only requirement is that the negative slide has to come from the same tissue tested.
What I used to do was to add buffer instead of the primary Ab and all the other reagents the same as in the slides run for specific Abs. You could also add non-immune globuline instead of the antibody but in this case you would need 1 negative control for each type Ab (1 negative for monoclonal Abs and 1 for polyclonal Abs).
I hope this will help you.
René J.
cynthia haynes <naje1972 <@t> yahoo.com> wrote:
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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