Intracellular Perforin and Granzyme B FCM

From: Paula Lavery (plaver@po-box.mcgill.ca)
Date: Wed Sep 01 1999 - 10:11:49 EST


I will be doing some intracellular staining for Perforin and Granzyme B on human leukocytes. Has anyone else done this before? Which antibodies did you use and at what concentrations? Was there anything special about the permeabilization protocol? (I'm already using the p-formaldehyde/methanol method for other intracellular stains.) Do I need to treat the cells in any way to prevent the molecules of interest from "leaking out" once the cells are permeabilized? Any and all suggestions would be appreciated. Thanks in advance!

_____________________________________________________
Paula Lavery
Transplant Immunology Laboratory
McGill University Health Centre
Royal Victoria Hospital
Montreal, Quebec, Canada
plaver@po-box.mcgill.ca

"If we knew what we were doing it wouldn't be called research, would it?"
-- Albert Einstein 



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