Recombinant Enhanced Green Fluorecence Protein

Synonyms eGFP

Accession P42212

GeneID 7011691

Source Escherichia coli.

Molecular Weight Approximately 26.9 kDa, a single non-glycosylated polypeptide chain containing 239 amino acids.

Quantity 10µg/50µg/1mg

AA Sequence MVSKGEELFT GVVPILVELD GDVNGHKFSV SGEGEGDATY GKLTLKFICT TGKLPVPWPT LVTTLTYGVQ CFSRYPDHMK QHDFFKSAMP EGYVQERTIF FKDDGNYKTR AEVKFEGDTL VNRIELKGID FKEDGNILGH KLEYNYNSHN VYIMADKQKN GIKVNFKIRH NIEDGSVQLA DHYQQNTPIG DGPVLLPDNH YLSTQSALSK DPNEKRDHMV LLEFVTAAGI TLGMDELYK

Purity > 95 % by SDS-PAGE and HPLC analyses.

Physical Appearance Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation Lyophilized from a 0.2 µm filtered concentrated solution in PBS, pH 7.4.

Endotoxin Less than 1 EU/µg of reGFP as determined by LAL method.

Reconstitution We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1 % BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions.

Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

- 12 months from date of receipt, -20 to -70 °C as supplied.

- 1 month, 2 to 8 °C under sterile conditions after reconstitution.

- 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Usage This material is offered by Shanghai PrimeGene Bio-Tech for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE.

SDS-PAGE

Reference 1. Prendergast FG, Mann KG. 1978. Biochemistry. 17:3448-53.

2. Shaner NC, Steinbach PA, Tsien RY. 2005. Nat Methods. 2:905-9.

3. Phillips GJ. 2001. FEMS Microbiol Lett. 204:9-18.

4. Wong FH, Banks DS, Abu-Arish A, et al. 2007. J Am Chem Soc. 129:10302-3.

Background Green fluorescent protein (GFP) here refers to the protein first purified from jellyfish Aequorea victoria, though many other organisms have similar proteins. It is a 26.9 kDa protein (composed of 238 a.a. residues) that shows green fluorescence in short-wave light (blue to ultraviolet). Despite of wild-type GFP, many mutants of GFP have been engineered for wider usage in research. Enhanced GFP (eGFP) has S65T and F64L mutations, which make GFP show increased fluorescence and fold more efficiently under 37℃, respectively. eGFP allows the use of GFP in mammalian cells. In A. Victoria, GFP plays roles as an energy transfer acceptor. It has long been used in cell and molecular biology as a reporter of gene expression. GFP can also been applied as a molecular thermometer to measure temperature accurately in fluids.