Hi Lisa:
  I used to run 1 negative control per block. The rationale was that you want to find out how the tissue itself reacts to those "non-antibody" components of the procedure.
  What I also did was to run 1 negative with "non-immune rabbit IgG" and another for mouse IgG (when I had polyclonal and monoclonal antibodies in the same run).
  Hope this will help you!
  René J.
lwuerges  wrote:
  I have a question regarding negative controls for a manual immunoperoxidase procedure. Do you use a neg control for each antibody used with each block? or one neg control per block regardless of how many antibodies for that block. The latter is what I was told, but the first seems more likely on account of each antibody may be treated differently with some needing antigen retrieval--obviously the contol should receive identical treatment. What is the usual neg control protocol?
Thank you.
Lisa Wuerges
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