[Histonet] Re: MOHS IPs

[Teri Johnson]

Hi Karen,

It could be that your cytokeratin antibody only recognizes a formalin fixed epitope conformation and enzyme digestion has nothing to do with it. Also try fixing in formalin and then try the stain again with and without the diluted enzyme digestion. Brief antigen retrieval (Citrate buffer pH 6.0, 30 minutes @ 60 degrees C) may also help. We have had good luck on minimally fixed frozen section staining using the lower temp AR with some antibodies.

Intermediate filaments are best preserved with alcohol-based fixatives. You could also try using 95% ethanol fixative for 5 minutes prior to staining, rinse with buffer and proceed with staining, no digestion.

Teri Johnson
Manager, Histology
Genomics Institute for
Novartis Research
Foundation
San Diego, CA
858-332-4752