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