Detection of GFP in supernatant - (Jul/06/2007 )

I have 293 cells that were transfected with a plasmid encoding GFP and a signal sequence for secretion. HOw can I detect the presence of GFP in the cell supernatant?

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I dont think that a flurometer is sensitive enough to pick up the signal. You may need WB or ELISA to do the job.

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highly concentrated GFP is to detect by appropriate excitation;

try to detect secretion into supernatant by a fluorescence plate reader for 488 nm; calibrate with a defined amount of GFP

GFP outside the cell is only poorly protected against oxidation; so, you may lose GFP signal after secretion;

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