Equine Gastrin EasyTest™ EIA Kit

Product Features

Product Name            Equine Gastrin 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                   18pg/mL
Assay Range                2.4-10000pg/mL
Reactivity                     Equine
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               GAST
Gene ID                         100034214
Swissprot ID               P55885

Standard Curve         Equine Gastrin EasyTest™ EIA Kit

Kit Components

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

Background

Gastrin is a peptide hormone that stimulates secretion of gastric acid by the parietal cells of the stomach and aids in gastric motility. It is released by G cells in the pyloric antrum of the stomach, duodenum, and the pancreas. Gastrin is found primarily in three forms: gastrin-34, gastrin-17 , gastrin-14.

Gastrin binds to cholecystokinin B receptors to stimulate the release of histamines in enterochromaffin-like cells, and it induces the insertion of K+/H+ ATPase pumps into the apical membrane of parietal cells.

Along with the above-mentioned function, gastrin has been shown to have additional functions as well: stimulates parietal cell maturation and fundal growth, Causes chief cells to secrete pepsinogen, increases antral muscle mobility and promotes stomach contractions, strengthens antral contractions against the pylorus, and constricts the pyloric sphincter, which diminishes the rate of gastric emptying, plays a role in the relaxation of the ileocecal valve, induces pancreatic secretions and gallbladder emptying.

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      

Equine Gastrin EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS