antibody

Green Fluorescent Protein (GFP)

G8965-04G

100 ul

monoclonal

mouse

nonconjugated

7C34

Green Fluorescent Protein (GFP)

mouse

Mab

Antibodies

100 ul

7C34

IgG2a

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. Applications: Suitable for use in Western Blot and ELISA. Other applications not tested. Recommended Dilution: Western Blot: 1-2ug/ml using Chemiluminescence technique. Antibodies react with native and denatured GFP or GFP containing proteins. Full length GFP is ~27-30kD. ELISA: 0.1-1ug/ml using 50-100ng control antigen/well. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile ddH2O or PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 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.

Full length GFP form jellyfish Aequorea victoria was expressed and purified (> 95%)

Recognizes wild type, recombinant, and enhanced form of GFP.

Purified using ion exchange and protein A/G chromatography.

Supplied as a lyophilized powder from PBS, 0.05% sodium azide.

As reported

Product Type: Mab Isotype: IgG2a Clone No: 7C34 Host: mouse Concentration: As reported Form: Supplied as a lyophilized powder from PBS, 0.05% sodium azide. Purity: Purified using ion exchange and protein A/G chromatography. Immunogen: Full length GFP form jellyfish Aequorea victoria was expressed and purified (> 95%) Specificity: Recognizes wild type, recombinant, and enhanced form of GFP. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

E WB

-20°C

Narayanan, S.R., J. Chromatogr., 658, 237 (1994); Olins, P.O., and Lee, S.C., Curr. Opin. Biotechnol., 4, 520 (1993); Uhlen, M., and Moks, T., Meth. Enzymol., 185, 129, (1990). Tsien, R.Y., Ann. Rev. Biochem., 67, 509 (1998); Chalifie, M., et al., Science, 263, 802 (1994); Cubitt, A.B., et al., Trends Biochem., 20, 448, (1995); Prasher, D.C., Trends Genet., 11, 320 (1995); Stearns, T., Curr. Biol., 5, 262 (1995); Errampalli, D., et al., J. Microbiol. Methods, 35, 187 (1999); Sawin, K.E., Methods Cell Biol., 58, 123 (1999); Cormack, B., Curr. Opin. Microbiol., 1, 406 (1998); Brand, A., Methods Cell Biol., 58, 165 (1999)