Rat Insulin ELISA Kit

Full Name: Rat Insulin ELISA Kit
Reactivity: Rat
Sample Type: Serum, Plasma
Sensitivity: 0.01 ng/ml

BACKGROUND

Insulin, a hormone produced in the pancreas by the beta cells of islets of Langerhans, has a molecular weight of 5800 Da and facilitates metabolism regulating carbohydrates. It plays an integral part in bodily functions. Insulin stimulates glucose-6-phosphatase which moves glucose into cells. When blood sugar levels rise, plasma insulin levels also increase. It also allows cells to uptake glucose that enters the bloodstream and uses it for energy. The hormone decreases blood sugar levels by moving sugar into cells using a cell membrane transporter called GLUT2, which is an acronym for “glucose transporter protein 2.” After the hormone’s effects are complete, it is then removed from the body by producing glycogen molecules in liver and muscle tissue to store excess sugar in the form of storage molecules. It is also known to possess a wide spectrum of biological actions and these include; increasing DNA replication and protein synthesis, controlling the cellular intake of glucose in adipose tissue and muscles, fatty acid synthesis, allosteric effect on numerous enzyme, gluconeogenesis, decreased proteinolysis, amino acid uptake, lipolysis and increased glycogen.

Plasma glucose levels have a major influence on insulin release and hyperglycemia is known to cause an immediate rise in blood levels of insulin. The neurological, metabolic and hormonal (glucagon, amino acids, and gastrointestinal hormone) factors mentioned above can also have an impact on the amount of insulin released. The subsequent assay will enable quantification of the insulin levels that are present, assisting in determining the correct amount needed to repair the shortage.

INTENDED USE

Rat insulin ELISA kit can be used for quantitative analysis of insulin levels using rat serum or plasma. This assay has a minimum sensitivity detection limit of 0.01 ng/ml.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

Coated Plate: Mouse monoclonal anti-insulin.
Calibrators (1-5): Concentration ranging between 0.156 – 10 ng/ml.
Calibrator 0.
Enzyme Conjugate.
Enzyme Conjugate Buffer.
Wash Buffer.
Substrate TMB.
Stop Solution.

SENSITIVITY

The standard range of insulin using current rat insulin ELISA kit was approximately 0.01 ng/ml. The dynamic range for this assay is 0.156 – 10 ng/ml.

ASSAY CHARACTERISTICS

– Detection limit: ≤0.15 μg/l
– Hook effect: Concentration up to at least 450 μg/l can be measured without giving falsely low results.
– Intra-assay-Variation: 3.2 – 10.0%
– Inter-assay-Variation: 2.8 – 5.1%
– Specificity: Human insulin (167%), Human proinsulin (75%), Human C-peptide (< 0.05%), Insulin lispro (Humalog® ) (167%), IGF-I (< 0.02%), IGF-II (< 0.02%), Rat C-peptide (< 0.001%), Rat proinsulin (7%), Mouse insulin (75%), Porcine Insulin (476%), Sheep Insulin (179%), Bovine Insulin (78%).

REFERENCES

A quantitative enzyme-linked immunosorbent assay for rat insulin. J Immunoassay. (1988) 9 (2): 135-58. Bank H.L.
Insulin therapy normalizes reduced myocardial β-adrenoceptors at both the onset and after sustained hyperglycemia in diabetic rats. Life Sci. (2015) 132: 101-7. Haley J.M., et al.
Development of an ultrasensitive immunoassay using affinity maturated antibodies for the measurement of rodent insulin. Anal Biochem. (2015) 473: 72-9. Imai S., et al.
Long-term treatment with atrial natriuretic peptide inhibits ATP production and insulin secretion in rat pancreatic islets. Am J Physiol Endocrinol Metab. (2011) 300 (3): E435-44. You H. and Laychock S.G.

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