Epidermal Growth Factor, Human (EGF), BioAssay(TM) ELISA Kit

Epidermal growth factor (EGF), a polypeptide mitogen, was first observed in 1959 by Cohen and Levi-Montalcini while studying Nerve Growth Factor (NGF) in snake venom extracts. It was subsequently isolated and purified from mouse submandibular glands. When injected into new-born mice this new factor caused precocious eyelid opening and incisor eruption.2,3 EGF was further purified, based on its ability to induce the proliferation of basal skin cells. Also, a potent inhibitor of gastric acid secretion was identified and isolated from the urine of a pregnant women, and named human ß-urogastrone. It was shown that this protein was very similar to purified human EGF.5,6, This EGF enzyme-linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal specific for EGF. Standards or samples are then added to the appropriate microtiter plate wells and incubated. EGF if present, will bind and become immobilized by the antibody pre-coated on the wells. The microtiter plate wells are thoroughly washed to remove unbound EGF and other components of sample. , In order to quantitate the amount of EGF present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated polyclonal antibody specific for EGF is added to each well to "sandwich" the EGF immobilized during the first incubation. The microtiter plate then undergoes a second incubation. The wells are thoroughly washed to remove all unbound HRP-conjugated antibodies and a TMB (3,3'5,5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain EGF and enzyme-conjugated antibody will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 2nm. In order to measure the concentration of EGF in the samples, this kit contains two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant/ urine testing). According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D.) versus EGF concentration (pg/ml). The concentration of EGF in the samples is then determined by comparing the O.D. of the samples to the standard curve. , Sensitivity: , Calibrator Diluent I: 20pg/ml, Calibrator Diluent II: 15pg/ml, Range: 31.3-1000pg/ml , Kit Components: , E3374-01A: Microtiter Plate 1x96wells , E3374-01B:. EGF (HRP) 1x15ml , *E3374-01C: EGF Standard 2x4ng, E3374-01D: Calibrator Diluent I (serum/plasma) 1x22ml , E3374-01E: Calibrator Diluent II (cell culture supernatant/urine) 1x22ml, E3374-01F: Wash Buffer (20X) 1x60ml , E3374-01G: Substrate A 1x11ml , E3374-01H: . Substrate B 1x11ml , E3374-01J: Stop Solution, H2SO4, 1x14ml , Storage and Stability: Store *E3374-01C powder at 4°C liquid at -20°C. Store at other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.