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Problem with double staining for FACS


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#1 Pham Dinh Chuong

Pham Dinh Chuong

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Posted 14 May 2009 - 02:51 AM

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.

#2 Hairpin

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Posted 15 May 2009 - 01:23 AM

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.


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




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