Double Staining - (Nov/07/2000 )

Hi! I have two questions;1.What protocol do you recommend to do double staining for two monoclonal antibodies? 2. If I use DAB and True Blue staining for both peroxidase ICC, how do you know the second substrate (True Blue) doens't react with peroxidase which use for detectection of the first primary antigen?Thank you----------------------------Akiko Iwata, Ph.DDivision of Cardiothoracic Surgery1959 N.E. Pacific Street, Room AA-115Box 356310Seattle, WA 98195-6310(206) 616-9847 (office)(206) 543-0325 (fax)

Heat slide in MW oven after first run. Heating denatures  antibodies in first run, and inactivates peroxidase.
see http://immunohistochemistry.onweb.cx