Hello:
I am trying to do IHC with rabbit anti human CDK4 (D-22) from Santa Cruz on
salivary gland tissue.  I am having background problems which is leading to
difficulty in figuring out what is positive and what is negative.  I have
used LSAB2 kit from Dako,  Rabbit Envision Polymer from Dako and the Rabbit
ABC Elite kit from Vector. I do a 3%  hydrogen peroxide treatment for 20
minutes and do antigen retrieval with 10mM Citric Buffer pH 6.0 for 45
minutes in a steamer and a 20 minute cool down.  I do a blocking step and
have used either serum free or goat serum for a block.

I already checked with the company about my protocol and it is the same as
theirs.  I tried doing it on skin which is reccommended as a control but
still have background problems.

Thanks
Joie Haviland
Senior Research Assistant
MD Anderson Cancer Research Center




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