Introduction
Gonadoliberin-1, also known as luteinizing hormone-releasing hormone (LH-RH) or gonadotropin-releasing hormone I (GnRH-I), is a peptide hormone that consists of 10 amino acids. Gonadoliberin-1 is produced by the hypothalamus in the brain. It is responsible for regulating the release of two other hormones, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), from the pituitary gland. Gonadoliberin-1 stimulates the production and release of LH and FSH, which are both involved in the reproductive system. LH stimulates the production of testosterone in males and ovulation and progesterone production in females, while FSH stimulates the growth and development of eggs in females and sperm production in males. Gonadoliberin-1 is regulated by various factors, including sex steroids and stress. In certain medical conditions, Gonadoliberin-1 analogs are used to regulate the levels of LH and FSH in the body.
The RayBiotech Gonadoliberin-1 Enzyme Immunoassay (EIA) Kit is an in vitro quantitative assay for detecting Gonadoliberin-1 peptide based on the competitive enzyme immunoassay principle.
In this assay, a biotinylated Gonadoliberin-1 peptide is spiked into the samples and standards. The samples and standards are then added to the plate, where the biotinylated Gonadoliberin-1 peptide competes with endogenous (unlabeled) Gonadoliberin-1 for binding to the anti-Gonadoliberin-1 antibody. After a wash step, any bound biotinylated Gonadoliberin-1 then interacts with horseradish peroxidase (HRP)-streptavidin, which catalyzes a color development reaction. The intensity of the colorimetric signal is directly proportional to the amount of captured biotinylated Gonadoliberin-1 peptide and inversely proportional to the amount of endogenous Gonadoliberin-1 in the standard or samples. A standard curve of known concentration of Gonadoliberin-1 peptide can be established and the concentration of Gonadoliberin-1 peptide in the samples can be calculated accordingly.
Product Features
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- HRP-Streptavidin
- TMB One-Step Substrate
- Stop Solution
- Assay Diagram
- Positive Control Sample
- Capture Antibody
- User Manual
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Orbital shaker
- Aluminum foil
- Saran Wrap
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- SigmaPlot software (or other software that can perform four-parameter logistic regression models)
Protocol Outline
- Prepare all reagents, samples and standards as instructed.
- Add 100 µl detection antibody to each well.
- Incubate 1.5 h at RT or O/N at 4°C.
- Add 100 µl standard or sample to each well.
- Incubate 2.5 h at RT.
- Add 100 µl prepared streptavidin solution.
- Incubate 45 min at RT.
- Add 100 µl TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl Stop Solution to each well.
- Read plate at 450 nm immediately.