This hGH 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 antibody specific to hGH. Standards or samples are then added to the microtiter plate wells and incubated. After wash all wells, hGH if present, will bind to the antibody pre-coated on the wells. In order to quantitatively determine the amount of hGH present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated monoclonal antibody specific to hGH is added to each well to “sandwich” the hGH immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, a TMB (3,3', 5,5' Tetramethyl-benzidene) substrate solution is added to each well. This enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain hGH and enzyme-conjugated antibody will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450 nm.

In order to measure the concentration of hGH in the sample; this Human hGH ELISA Kit includes a set of calibration standards (6 standards). The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density (O.D.) versus GH concentration (ng/mL). The concentration of hGH in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Platform

Microplate

Detection Type

Colorimetric

Sample Type

Serum

Sensitivity

< 0.5 ng/ml

Product Range

0.5-50 ng/ml

Reactivity

Human

Cross Reactivity

This assay exhibits no detectable cross-reaction with LH, hCG, TSH, Prolactin, and FSH.

Recovery

Storage

Store at 2-8°C.

Synonyms

gH, GH-N, GH1, GHN, Growth hormone, Growth hormone 1, HGH, Pituitary growth hormone, Somatotropin, SOMA_HUMAN

Disclaimer

This product is for research use only. It is not intended for diagnostic or therapeutic use.

Scientific Validation Data (1)

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Figure 1: Standard Curve - Human Growth Hormone ELISA Kit (A33015)

Representative Standard Curve using Human Growth Hormone ELISA Kit (EL10030).

Citations (1)

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(3)

Magneto-Immunoassay for the Detection and Quantification of Human Growth Hormone

References: Human Growth Hormone ELISA Kit (A33015)

Abstract:

Physiological and endocrine maintenance of a normal human growth hormone (hGH) concentration is crucial for growth, development, and a number of essential biological processes. In this study, we describe the preparation and characterization of magnetic nanoparticles coated with a gold shell (MNPs-Au). The optimal surface concentration of monoclonal anti-hGH antibodies (m-anti-hGH) on magnetic nanoparticles, as well as conditions that decrease non-specific interactions during the magneto-immunoassay, were elaborated. After the selective recognition, separation, and pre-concentration of hGH by MNPs-Au/m-anti-hGH and the hGH interaction with specific polyclonal biotin-labeled antibodies (p-anti-hHG-B) and streptavidin modified horseradish peroxidase (S-HRP), the MNPs-Au/m-anti-hGH/hGH/p-anti-hGH-B/S-HRP immunoconjugate was formed. The concentration of hGH was determined after the addition of 3,3',5,5'-tetramethylbenzidine and hydrogen peroxide substrate solution for HRP; the absorbance at 450 nm was registered after the addition of STOP solution. The developed sandwich-type colorimetric magneto-immunoassay is characterized by a clinically relevant linear range (from 0.1 to 5.0 nmol L^-1, R² 0.9831), low limit of detection (0.082 nmol L^-1), and negligible non-specific binding of other antibodies or S-HRP. The obtained results demonstrate the applicability of the developed magneto-immunoassay for the concentration and determination of hGH in the serum. Additionally, important technical solutions for the development of the sandwich-type colorimetric magneto-immunoassay are discussed.

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