Mouse Monoclonal Ki-67 ? - (Jan/30/2006 )

Could anyone help me with suggestions for optimal conditions for the antigen retrieval step for Ki-67 in mouse large intestine for a IHC assay? I am trying to stay away from heat induced epitope retrieval (HIER) and would like to use a proteolytic induced epitope retrieval (PIER) specifically using proteinase K in my assay. Any optimal conditions using proteinase K or other Proteolytic methods would be appreciated.

I use Pepsin preheated to 37C, after rehydration and peroxidase blocking , 200ul of pepsin at 37c for 10 min. Just becarful during wash since few slides tend to lift over.

I also use 0.1% trypsin again preheated to 37C applied for 10-15min at 37C could give similar results. Zymed has a product called "Digest all" that has Ficin, trypsin, & Pepsin.

Good luck