Description
The Glucagon-HS ELISA, developed and manufactured in Japan by Yanaihara Institute Inc., is used for quantitative determination of glucagon in plasma, serum and culture medium supernatant. The kit is characterized by its ultrasensitive quantification and high specificity. In addition, it has no influence by other components in samples. Glucagon standard is highly purified synthetic product. The kit is validated for human, mouse, and rat samples.
Glucagon is a 29-amino acid polypeptide hormone, synthesized and secreted from alpha cells of the islets of Langerhans. Glucagon generally elevates the concentration of glucose in the blood by promoting gluconeogenesis and glycogenolysis, and it has the opposite effect of insulin. The secretion of glucagon and insulin into the blood in response to the blood glucose concentration is the primary mechanism responsible for keeping the glucose levels.
Principle of Method
This ELISA kit for determination of glucagon is based on a sandwich enzyme immunoassay with two monoclonal antibodies. Standards or samples, and HRP labeled antibodies are added to the wells of plate coated with antibodies against glucagon. During the incubation antibody – antigen – labeled antibody complex is formed on the surface of the wells. After the incubation and rinsing out excess labeled antibody, HRP enzyme activity is finally determined by 3,3’,5,5’-Tetramethylbenzidine (TMB) and the concentration of glucagon is calculated.
Specifications
| Method | One-step sandwich ELISA |
|
Sample Type |
Plasma (EDTA-2Na), serum, cell culture medium supernatant |
|
Sample Volume |
10 μL |
| Assay Time | 20 - 24 hours + 0.5 hour |
| Assay Range | 2.2 - 143.6 pmol/L (7.8 - 500 pg/mL) |
| Sensitivity | 0.3 pmol/L (1.08 pg/mL) |
| Antibodies | Monoclonal |
| Reactivity | Human, mouse, rat |
| Storage | 2-8ºC |
| Manufacturer | Yanaihara Institute |
| Country of Origin | Japan |
Standard Curve
Specificity
This ELISA kit has high specificity to glucagon, and shows no significant cross reactivity to Glicentin, 3 Oxyntomodulin, GLP-1 and GLP-2.
Precision & Reproducibility
| Test sample | Intra-assay CV (%) | Inter-assay CV (%) |
| Serum (Human, Rat, Mouse) | 1.8~ 3.5 | 3.5~ 9.1 |
| Plasma (Human, Rat, Mouse) | 2.1~4.6 | 3.4~7.1 |
Stability and Storage
Store all of the components at 2-8ºC. The kit is stable under the condition for 21 months from the date of manufacturing. The expiry date is stated on the label of kit
Resources
References
- Unger, R.H., Eisentraut, A.M., McCall, M.S., Keller, S., Lanz, H.C. and Madison, L.L. (1959)： Glucagon antibodies and their use for immunoassay for glucagon. Proc．Soc．Exp．Biol．Med.,102： 621 - 623 2.
- Unger, R.H., Eisentraut, A.M., McCall, M.S. and Madison, L.L. (1961)： Glucagon antibodies and an immunoassay for glucagon. J．Clin．Invest.,40：1280-1289 3.
- Nishino, T., Kodaira, T., Shin, S., Imagawa, K., Shima, K., Kumahara, Y., Yanaihara, C. and Yanaihara, N.（1981）： Glucagon radioimmunoassay with use of antiserum to glucagon C-terminal fragment. Clin．Chem.,27：1690-1697 4.
- Jaspan, J.B., and Rubenstein, A.H. (1977)： Cerculating glucagon：Plasma profiles and matabolism in health and disease. Diabetes., 26：887-902 5.
- Glucagon related peptides (1993)： (Okuno, G., Ohneta, A. and Shima, K.ed.) pp．52-65．Ishiyaku, Tokyo
Ilex Life Sciences LLC is an authorized distributor of Yanaihara Glucagon-HS ELISA.