we're running one negative normal serum slide per block. For example, if we're performing mouse antibodies i.e. CD-45, HMB-45, pan Cytokeratin, we run one IgG mouse normal serum control. If we are running rabbit polyclonals (or rabbit monoclonals) and mouse monoclonals on the same case, we usually would run a IgG normal mouse and normal rabbit serum slide. Now, I know the purists out there are going to tell me what if I use an IgM mouse monoclonal, I should be running an IgM mouse normal serum. I agree, but my budget doesn't allow to run a negative for all scenarios. In that frame of mind, if we run a case with multiple different pretreatments, we run a negative on the harshest pretreatment. Again, not ideal, but I think it is better than just putting PBS or TBS buffer on a slide and calling that negative. That's my 3 cents worth and I'm sticking to that story.

JTT
----- Original Message ----- From: "Sheila Haas" <micropathl...@yahoo.com>
To: <histonet@lists.utsouthwestern.edu>
Sent: Wednesday, April 08, 2009 1:57 PM
Subject: [Histonet] Negative IHC controls


I have a question concerning ANP.22570 on the CAP checklist. Could you all tell me how you are handling negative controls for IHC staining? The question actually states (and I've confirmed with CAP) that we should be running two types of negative controls. One for reagents and one for each antibody in a run. I'd like to know what the practice is. This seems very costly and time consuming. Thanks in advance!

Sheila Haas
Laboratory Supervisor
Micro Path Laboratories



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