Epidermal Growth Factor (EGF) was originally discovered in crude preparations of nerve growth factor prepared from mouse submaxillary glands. Its actions were shown to induce early eyelid opening, incisor eruption, hair growth inhibition, and stunt growth when injected into newborn mice. Human EGF was isolated from urine based on its inhibitory effect on gastric secretion and named urogastrone, accordingly. EGF is prototypic of a family of growth factors that are derived from membrane-anchored precursors. All members of this family are characterized by the presence of at least one EGF structural unit (defined by the presence of a conserved 6 cysteine motif that forms three disulfide bonds) in their extracellular domain. EGF is initially synthesized as a 130kD precursor transmembrane protein containing 9 EGF units. The mature soluble EGF sequence corresponds to the EGF unit located proximal to the transmembrane domain. The membrane EGF precursor is capable of binding to the EGF receptor and was reported to be biologically active.
Sample Type
Cell culture supernatants, serum, plasma, and tissue
Intended Use
The Human EGF BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human EGF concentrations in cell culture supernatants, serum, plasma, and tissue.
Specificity
Recognizes human EGF.
Test Principle
This assay employs the Sandwich immunoassay technique. An anti-h EGF monoclonal coating antibody is adsorbed onto microwells. EGF present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h EGF antibody is added and binds to EGF captured by the first antibody. Following incubation unbound Biotinylated anti-h EGF antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h EGF antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of EGF present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven EGF standard dilutions and EGF sample concentration determined.
Kit Components
Aluminium pouches with a Microwell Plate: 8x12Standard: 2xVialBiotinylated antibody: 2xVialStreptavidin-HRP: 2xVialAssay Buffer (25ml/Vial): 2xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x12mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 3xstripsInstruction: 1x
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
