Green fluorescence protein (GFP) is a 27kD protein derived from the bioluminiscent jellyfish Aquorea victoria, emiting green light (lambda=509 nm) when excited (excitation by Blue or UV light, absorption peak lambda=395 nm). GFP is a useful tool in cell biology research, as its intrinsic fluorescence can be visualized in living cells. Light-stimulated GFP fluorescence is species-independent and a fluorescence has been reported from many different types of GFP-expressing hosts, including microbes, invertebrates, vertebrates and plants. No exogenous substrates and cofactors are required for the fluorescence of GFP, since GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP is widely used as a reporter (tag) for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without any further staining. Other applications of GFP include measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. To increase a fluorescence intensity of GFP, chomophore mutations have been created. The Enhanced GFP has a fluorescence 35 times more intense than the wt-GFP. Mutagenesis of GFP has produced also many mutants (e.g. yellow fluorescent protein, cyan fluorescent protein) with warying spectral properties. Antibodies raised against full-length GFP variants should also detect other variants of the protein. Applications: Suitable for use in Immunoprecipitation, Flow Cytometry and Western Blot. Other applications not tested. Recommended Dilutions: Western Blot: 0.5-1.5ug/ml, positive control: transfected cells, negative control: non-transfected cells. Immunocytochemistry: 1-3ug/ml Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
