F4/80 thanks it's working great!
Many, many thanks to all who replied to my request for help. Special thanks to
Barbara Wright for providing me with her SOP which gave me a great starting place.
Also, Barbara, I liked the format of your SOP so much that I slightly modified it
and changed over all of our IHC SOP sheets.
First I bought the antibody from Serotec cat# MCAP497 (never could get the Accurate
Sci. antibody to work), dilution 1:100, 45 minutes at 30oC
I also used Serotec's STAR72 goat anti rat, mouse adsorbed, HRP conjugated at a
dilution of 1:50 for 15 minutes at 30oC
Because I was in a hurry as I wanted instant (OK a couple of hours) gratification so
instead of an over night incubation at 4 oC with no pretreatment I did a short
pre-treatment with trypsin for 7 minutes at 37oC. The results are beautiful.
Thanks again,
Bob
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
University of North Carolina at Chapel Hill
919-966-6343
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