Problem with double staining for FACS - (May/14/2009 )
My protein can uptake into cells as a single agent and it can inhibit the replication of virus. First, i treat protein, then infect virus. I stain sample as double staining with FITC and PE. But i can only detect virus, not my protein. When i detect my protein using only PE, i can see the uptaken protein.
Would you please give me some advices? Thanks.
-Pham Dinh Chuong-
Pham Dinh Chuong on May 14 2009, 12:51 PM said:
My protein can uptake into cells as a single agent and it can inhibit the replication of virus. First, i treat protein, then infect virus. I stain sample as double staining with FITC and PE. But i can only detect virus, not my protein. When i detect my protein using only PE, i can see the uptaken protein.
Would you please give me some advices? Thanks.
Would you please give me some advices? Thanks.
Hi 'Pham Dinh Chuong',
I need a little bit more information. Do you stain your cells with a PE-coupled antibody against your Protein and a FITC- conjugated antibody against the virus? How do you permeabilize and fix your cells?
I suppose you work in a cell culture system, or?
Do you use a color-compensation?
Regards
-Hairpin-