Hi
I am trying to demonstrate T cells using Polyclonal Rabbit Anti human CD3, T cell (DAKO ENVISION).  The control tissue is tonsil and using a manual procedure.  I have increased the time in the microwave pressure cooker from 5 up to 30 minutes (pH 6 buffer) and have no success.  The antibody is incubated for 30 minutes (recommended) but I have gone as high as 1 hour.  I am using AEC and Mayers counterstain.  Any help or advise would be greatly appreciated.
				Diana 

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