Description
The Total GLP-1-HS ELISA kit, developed and manufactured in Japan by Yanaihara Institute Inc., is used for quantitative determination of total GLP-1 in plasma or cell culture supernatant samples. The kit can be used with mouse, rat, or human samples. The kit is characterized by sensitive quantification and high specificity. In addition, it is not influenced by other components in plasma sample and no sample pre-treatment is required.
Summary
GLP-1 is a peptide hormone from the intestinal mucosa, which is produced from its precursor, proglucagon by post transnational processing. The mammalian proglucagon is synthesized in the neuroendocrine L-cell of the intestine and the alpha-cells of the pancreas. It contains within its structure the sequences of glucagon and two glucagon-like peptides (GLP-1 and GLP-2) in tandem flanked at their amino and carboxyl termini by dibasic residues. GLP-1 is a 37 amino acids peptide and produced in the small intestine and in the pancreas in the human, in either C-terminal-amidated on glycine-extended form.
Principle of Method
This kit for determination of total GLP-1 in mouse, rat and human plasma and culture supernatant sample is based on the sandwich enzyme immunoassay. During first immune incubation, GLP-1(7-36)amide in standards or in samples bind to the rabbit anti GLP-1(7-36)amide antibody, which is coated on the surface of the microtiter plate. After incubation and plate washing, labeled antibody solution (biotinylated rabbit anti GLP-1(7-36)amide polyclonal antibody) is added to bind to the antibody-antigen complex. Then, HRP labeled streptavidin (SA-HRP) is added to form antibody-antigen-biotinylated antibody complex. Finally, HRP enzyme activity is determined by 3,3’,5,5’-Tetramethylbenzidine (TMB) and the concentration of total GLP-1 is calculated.
Specifications
| Method | Sandwich ELISA |
|
Sample Type |
Plasma (EDTA-2Na), cell culture medium supernatant |
|
Sample Volume |
25 μL (10 μL for mouse) |
| Assay Time | 18 + 1 + 0.5 and 0.5 hr |
| Assay Range | 1.24 – 300 pM (4.07 - 989.31 pg/mL)) |
| Antibodies | Polyclonal |
| Reactivity | Rat, mouse, human |
| Storage | 2-8ºC |
| Manufacturer | Yanaihara Institute |
|
Country of Origin |
Japan |
Standard Curve
Specificity
This ELISA kit has high specificity to GLP-1(1-36) amide, GLP-1(7-36) amide and GLP-1(9-36) amide and shows low crossreactivity to GLP-1(1-37) and GLP-1(7-37), and shows no crossreactivity to rat and human GLP-2, rat Glicentin, human Glucagon, rat mouse and human GIP (1-42), rat and human GIP (3-42).
Precision & Reproducibility
| Test sample | Intra-assay CV (%) | Inter-assay CV (%) |
| Human Plasma |
1.98 – 5.43 | 2.21 – 3.84 |
| Mouse Plasma | 4.87 – 4.98 | 4.86 – 9.76 |
Stability and Storage
Store all of the components at 2-8ºC. The kit is stable under the condition for 24 months from the date of manufacturing. The expiry date is stated on the label of kit.
Resources
Ilex Life Sciences LLC is an authorized distributor of Yanaihara Total GLP-1-HS ELISA.