How is GFP detected?
Posted November 14, 2022
GFP is detected through conventional techniques such as fluorescent microscopy and flow cytometry. Flow cytometry is a sensitive and effective method to quantitatively analyze fluorescent intensity. Through marking expression viral vectors that contain gene products with GFP, the expressed vectors in this population can be identified and used for reconstitution of organisms. Fluorescent microscopy is used to identify the subcellular location and expression of GFP. Proteins are tagged with GFP through genetic modification in order to be analyzed through fluorescent microscopy. GFP acts as a fluorescent protein reporter by cloning GFP with the target protein at the N- or C- terminus. GFP expression may also be detected by fluorescence-activated cell sorting (FACS) analysis or fluorometer assays 24-72 hours post transfection depending on the cell line being used for cell culture. FACS is a rapid technique for isolating large numbers of fluorescently tagged cells from a heterogeneous mixture of cells.