Rat VIP EasyTest™ EIA Kit

Product Features

Product Name            Rat VIP 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                   0.5pg/mL
Assay Range                2.4-10000pg/mL
Reactivity                     Rat
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               VIP; vip/phi27
Gene ID                        117064
Swissprot ID              P01283

Standard Curve       Rat VIP EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-VIP Antibody
• Standard VIP peptide
• Biotinylated VIP 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-VIP 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 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, VIP binds with big proteins. Acidification of blood is required to separate and purify VIP.

Background

Vasoactive intestinal peptide also known as VIP is a peptide hormone containing 28 amino acid residues. VIP is neuropeptide that belongs to a glucagon/secretin superfamily, the ligand of class II G protein-coupled receptors. VIP is produced in many tissues of vertebrates including the gut, pancreas, and suprachiasmatic nuclei of the hypothalamus in the brain.

With respect to the digestive system, VIP seems to induce smooth muscle relaxation, stimulate secretion of water into pancreatic juice and bile, and cause inhibition of gastric acid secretion and absorption from the intestinal lumen. Its role in the intestine is to greatly stimulate secretion of water and electrolytes, as well as stimulating contraction of enteric smooth muscle, dilating peripheral blood vessels, stimulating pancreatic bicarbonate secretion, and inhibiting gastrin stimulated gastric acid secretion. These effects work together to increase motility. It also has the function of stimulating pepsinogen secretion by chief cells. VIP helps to regulate prolactin secretion.

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      

Rat VIP EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS