Human Vasopressin EasyTest™ EIA Kit

Product Features

Product Name            Human Vasopressin EasyTest™ EIA Kit
Assay Type                   Solid Phase Competitive ELISA
Format                           96-well strip plate
Assay Process             Two wash step, three incubations
Assay Length              2.0 hours
Sample Volume         Serum , Plasma, 50µL
Detection Method    Colorimetric
Sensitivity                   7pg/mL
Assay Range                2.4-10000pg/mL
Reactivity                     Human
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               AVP; ADH; ARVP-NPII; AVRP; VP
Gene ID                         551
Swissprot ID               P01185

Standard Curve        Human Vasopressin EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-Vasopressin Antibody
• Standard Vasopressin peptide
• Biotinylated Vasopressin peptide
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 25 µl of anti-Vasopressin antibody to all wells.
3. Add 75 µl of 1 x assay diluent to the blank wells and 50 µl to B0 wells.
4. Add 50 µl of standard peptide/samples to the appropriate wells.
5. Add 25 µl of biotinylated peptide into each well except the blank wells.
6. Incubate 1.0 h at RT.
7. Wash and add 100 µl of diluted streptavidin-HRP solution to all wells.
8. Incubate 45 min at RT.
9. Wash and add 100 µl of TMB Substrate Solution into each well.
10. Incubate 15 min at RT.
11. Add 50 µl of Stop Solution to each well.
12. Read at 450 nm/570 nm.

Application

In blood, vasopressin binds with big proteins. Acidification of blood is required to separate and purify vasopressin.

Background

Vasopressin, also known as arginine vasopressin (AVP), antidiuretic hormone (ADH), is aneurohypophysial hormone found in most mammals. It is derived from a preprohormone precursor that is synthesized in the hypothalamus and stored in vesicles at the posterior pituitary. Most of it is stored in the posterior pituitary to be released into the bloodstream. However, some AVP may also be released directly into the brain, and plays an important role in social behavior, sexual motivation and pair bonding, and maternal responses to stress.

The vasopressins are peptides consisting of nine amino acids. The amino acid sequence of arginine vasopressin is Cys-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly, with the cysteine residues forming a disulfide bond. Lysine vasopressin has a lysine in place of the arginine. Its two primary functions are to retain water in the body and to constrict blood vessels. Vasopressin regulates the body’s retention of water by acting to increase water reabsorption in the collecting ducts of the kidney nephron. Vasopressin is a peptide hormone that increases water permeability of the kidney’s collecting duct and distal convoluted tubule by inducing translocation of aquaporin-CD water channels in the kidney nephron collecting duct plasma membrane. It also increases peripheral vascular resistance, which in turn increases arterial blood pressure.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Human Vasopressin EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS