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

£943.00 96 Tests

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.1 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.4 Also, a potent inhibitor of gastric acid secretion was identified and isolated from the urine of a pregnant women, and named human ?-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.

SKU: USE3374-01 Categories: ,

Additional information

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.

Shipping Temperature

On Dry Ice

Source Antigen

Human

Shelf Life

6 Months