Green Fluorescent Protein, ELISA Control (GFP)

Recombinant DNA technology allows the addition of short pieces of well-defined tags, "peptides" or proteins at the amino or c-terminus of target genes, which can provide 'affinity handles' designed to bind specific matrices. Therefore, tags enables a selective identification and purification of the protein of interest. The addition of a green fluorescent protein (GFP) tag to a given gene, creates a stable fusion product that does not appear to interfere with the bioactivity of the protein, or with the biodistribution of the GFP tagged product. GFP is a 27 kD (238 a.a.) protein, derived from the bioluminescent jellyfish Aequorea victoria, in which light is produced when energy is transferred from the Ca2+-activated photoprotein aequorin to GFP. GFP is acknowledged as a unique tool to monitor dynamic processes in a variety of living cells or organisms. When expressed in either eukaryotic or prokaryotic cells and illuminated by blue or UV light, GFP yields a bright green fluorescence. Light-stimulated GFP fluorescence is species-independent and a fluorescence has been reported from many different types of GFP-expressing hosts, including microbes, invertebrates, vertebratesand plants. Exogenous substrates and cofactors are not required for the fluorescence of GFP, since GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein. Additionally, detection of GFP and its variants can be performed with living tissues instead of fixed samples. GFP signals can be quantified by flow cytometry, confocal scanning laser microscopy, and fluorometric assays. Indeed, many recombinant proteins have been engineered with GFP tags to facilitate the detection, isolation and purification of the proteins. The potential applications have been multiplied by the introduction of brighter GFP mutants and mutants with modified spectral properties, like the blue fluorescent protein (BFP), which allow the independent detection of BFP- and GFP- tagged proteins, even when coexpressed in the same cell. Monoclonal antibody reacting specifically with GFP may be useful in various immunotechniques,to identify the expression of a GFP fusion protein in situ and by immunoblotting, in bacteria, bacterial lysates or cells and tissues transfected with a GFP fusion protein expressing vectors. It may also be used to correlate levels of GFP protein expression with fluorescence intensity and for immunoprecipitation of GFP fusion proteins. Source: Full length GFP protein (238aa, ~27kD) form jellyfish aequorea Victoria was expressed in E. coli and purified (> 95%). Applications: Suitable for ELISA coating or for standards or FACS control. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer., ,