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Insulin ELISA Kit

INTENDED USE

Human insulin ELISA kit is a reliable quantitative procedure for measuring insulin in human serum and plasma. This assay has a minimum sensitivity limit of 1.76 µIU/ml.

BACKGROUND

Insulin is a polypeptide hormone that is secreted from the pancreas by beta cells of the islets of Langerhans, it has a molecular weight of 5800 Da and plays an important role in regulating carbohydrate metabolism. Insulin 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. Insulin secretion can be mainly controlled by the levels of plasma glucose where hyperglycemia is known to induce a prompt increase in levels of insulin in circulation. Other factors which can also have an influencing factor include neural, metabolic and hormonal (glucagon, amino acids and gastrointestinal hormone) can all participate in controlling the levels of insulin that is being secreted.

The following assay will allow the determination of quantitative levels insulin that are present, thereby helping to determine what the correct dose of insulin is required in order to rectified the deficiency. There are some conditions where human insulin disturbance is pathologic such as metabolic syndrome, diabetes mellitus, polycystic ovary syndrome and insulinoma. Diabetes mellitus exists in two forms: type 1 where in the conditions of absolute insulin deficiency there is an autoimmune-mediated destruction of human insulin producing beta cells in the pancreas and type 2 where this form is strongly inherited which is the result of a combined influence of genetic susceptibility and environmental factors. In both of these cases there needs to be an increase in the production of insulin either by delivering insulin by intravenous or oral methods, or by taking medication.

INSULIN ELISA KIT 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.

  • Microtiterwell Plate: Wells coated with monoclonal anti insulin antibody.
  • Zero Standard: 0 µIU/mL.
  • Standard 1-5: Concentration 6.25, 12.5, 25, 50, 100 µIU/mL. Made up from anti human insulin. The standards are calibrated against international WHO approved Reference material NIBSC 66/304.
  • Enzyme Conjugate: Mouse monoclonal anti-insulin antibody conjugated to biotin.
  • Enzyme Complex: Streptavidin-HRP Complex.
  • Substrate Solution: Tetramethylbenzidine (TMB).
  • Stop Solution: 0.5 M H2SO4, Avoid contact with the stop solution. It may cause skin irritations and burns.
  • Wash Solution (40x Concentrated).

SENSITIVITY

The minimum sensitivity detection limit of human insulin using current insulin ELISA kit was 1.76 µIU/ml. The dynamic range for this assay is 6.25 – 100.0 µIU/ml.

ASSAY CHARACTERISTICS

– Insulin: ELISA
– Assay Dynamic Range: 6.25 – 100.0 µIU/ml
– Sensitivity: 1.76 µIU/ml
– Intra-Assay: 1.8 – 2.6%
– Inter-Assay: 2.9 – 6.0%
– Recovery: 91.8 – 109.6%
– Linearity: 88.4 – 110.4%
– Interference: None
– High Dose Effect: None up to 1600 µIU/ml of insulin.

REFERENCES

  1. Evolution of insulin: from human to analog. Am J Med. (2014) 127 (10 Suppl): S25-38. Review. Tibaldi J.M.
  2. Insulin Signaling in the Control of Glucose and Lipid Homeostasis. Handb Exp Pharmacol. (2016) 233: 51-71. Review. Saltiel A.R.
  3. Insulin tactics in type 2 diabetes. Med Clin North Am. (2015) 99 (1): 157-86. Review. Meah F. and Juneja R.

ADDITIONAL INFORMATION

  • Full Name: Insulin ELISA Kit
  • Reactivity: Human
  • Sample Type: Serum, Plasma
  • Sensitivity: 1.76 µIU/ml

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Your secretory IgA ELISA gave good results and I was also really impressed with how quickly we received it.

L. Johnston
PhD Student / University of Glasgow

It is refreshing to know that you have a technical team that is very knowledgeable. I have already recommended your company to other researchers in our department.

Dr. P. Anderson
Lecturer / University College London (UCL)

I am a first time user and found that your instruction manual was very easy to follow. The insulin ELISA assay performed well and I was happy with the results that were generated.

J. Thomas
Senior Technician / Addenbrooke’s Hospital

I carried out a pilot study comparing the performance of many ELISA assay's from different suppliers and found your kits to be one of the better performers. We observed good linearity and tight replicates.

Dr. C. Davies
Lead Scientists / AstraZeneca

You are my first point of contact when I am looking to purchase ELISA. You have such an easy and simple system, yet it is very effective.

A. Shaw
Purchasing / University of Oxford