We make up our own antibody cocktail of 34ßE12 (DAKO dilution 1:150) and P63
(DAKO 1:400) and stain the section with this cocktail with a Polymer
peroxidise detection system (DAB).(Heat retrieval in a high pH retrieval
solution for 20 min at 100 C.
The stained slides are washed and darkened using a copper sulphate solution.
Placed in a buffer wash and then stained for the second time with P504S
(AMACR/Racemase) at a dilution of 1:150 with a Polymer Alkaline Phosphatase
detection system (Fast Red).
We do this all on the BondMax Immunostainer.
We have been doing this for over 6 months now and did it this way because we
already had all the concentrated antibodies.
Proper fixation is vital.
Hope this helps.
Royal Brisbane Campus
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