Introduction
Neurokinin B is a peptide hormone belonging to the tachykinin family, which also includes Substance P and Neurokinin A. All the peptides from tachykinin families are derived from two preprotachykinin genes - the PPT-A gene and PPT-B gene. The former encodes Substance P. Neurokinin A & Neuropeptide K, and the latter encodes Neurokinin B.
Neurokinin B has shown important roles in regulating immune and neurological functions in humans. It is found in higher concentration in women suffering from pre-eclampsia during pregnancy. It is reported to stimulate the production of immunoglobulins in peripheral B lymphocytes via its NK-3 receptor. Recently Neurokinin B, along with its NK-3 receptor, has shown to play a critical role as a key neuroregulatory switcher on human puberty, governed by the brain through the release of the hormone GnRH, which starts a series of processes that ultimately leads to the production of sex hormones.
Neurokinin B has shown potential clinical application as a biomarker in pre-eclampsia during pregnancy and onset of puberty.
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.Storage/Stability
Standard, Biotinylated Neurokinin B peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.