Human GLP-1 EasyTest™ EIA Kit

Product Features

Product Name            Human GLP-1 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                   90pg/mL
Assay Range                24.4-100000pg/mL
Reactivity                     Human
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               GCG; GLP1; GLP2; GRPP
Gene ID                         2641
Swissprot ID               P01275

Standard Curve        Human GLP-1 EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-GLP-1 Antibody
• Standard GLP-1 peptide
• Biotinylated GLP-1 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-GLP-1 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, GLP-1 binds with big proteins. Acidification of blood is required to separate and purify GLP-1.

Background

Glucagon-like peptide-1 (GLP-1) is an incretin derived from the transcription product of the proglucagon gene. The biologically active forms of GLP-1 are: GLP-1-(7-37) and GLP-1-(7-36) NH2. Those peptides result from selective cleavage of the proglucagon molecule.

GLP-1 secretion by ileal L cells is dependent on the presence of nutrients in the lumen of the small intestine. The secretagogues of this hormone include major nutrients like carbohydrate, protein and lipid. Once in the circulation, GLP-1 has a half-life of less than 2 minutes, due to rapid degradation by the enzyme dipeptidyl peptidase-4. It is a potent antihyperglycemic hormone, inducing glucose-dependent stimulation of insulin secretion while suppressing glucagon secretion. GLP-1 appears to restore the glucose sensitivity of pancreatic β-cells, with the mechanism possibly involving the increased expression of GLUT2 and glucokinase. GLP-1 is also known to inhibit pancreatic β-cell apoptosis and stimulate the proliferation and differentiation of insulin-secreting β-cells. In addition, GLP-1 inhibits gastric secretion and motility. This delays and protracts carbohydrate absorption and contributes to a satiating effect.

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 GLP-1 EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS