The green fluorescent protein (GFP) is a protein composed of 238aa residues (26.9kD) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. In cell and molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms it has been used to make biosensors, and many animals have been created that express GFP as a proof-of-concept that a gene can be expressed throughout a given organism. The GFP gene can be introduced into organisms and maintained in their genome through breeding, injection with a viral vector, or cell transformation. To date, the GFP gene has been introduced and expressed in many Bacteria, Yeast and other Fungi, fish (such as zebrafish), plant, fly, and mammalian cells, including human. Sample Type: Cell lysates Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of GFP. Sensitivity: <10pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant GFP. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This GFP BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for GFP has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: S2-K238) and test samples are added to the wells, a biotinylated detection specific for GFP is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the All Animals GFP amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-GFP antibody, 1x plate Lyophilized recombinant GFP standard, 2x10ng/tube Biotinylated anti-GFP antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
