Cat No.: 32270 ★Download Datasheet★ ★MSDS★
Introduction
Insulin is a peptide hormone exclusively produced in pancreatic beta-cells. It consists of A chain and B chain, which are linked by two disulfide bridges. Insulin is the primary hormone responsible for glucose metabolism. Impaired insulin secretion and insulin
resistance are key causes of type 2 diabetes (T2D).
Principle of the Assay
This assay is a two-site ELISA. The micro-plate is pre-coated with a monoclonal antibody against insulin. Standards and samples are added into the wells and coincubated with a monoclonal antibody conjugated to horseradish peroxidase (HRP) enzyme. After wash
step to remove any unbound substances, TMB substrate is added and colour develops in proportion to the amount of insulin bound
initially. The assay is stopped and the optical density of the wells determined using a micro-plate reader. Since the increases in
absorbance are directly proportional to the amount of captured insulin, the unknown sample concentration can be interpolated from a reference curve included in each assay.
Assay Performance
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each assay.
Insulin (ng/mL) | Absorbance (450 nm) | Blanked Absorbance |
0 | 0.052 | 0 |
0.2 | 0.063 | 0.011 |
0.5 | 0.098 | 0.046 |
1.0 | 0.197 | 0.145 |
2.0 | 0.558 | 0.506 |
3.5 | 1.23 | 1.178 |
7 | 2.731 | 2.679 |
B. Sensitivity
The lowest insulin level that can be measured by this assay is 0.2 ng/mL.
C. Precision
Intra-assay Precision (Precision within an assay) C.V<6.1%.
Inter-assay Precision (Precision between assays) C.V<2.9%.
D. Recovery
The recovery of the assay was determined by adding various amounts insulin to a sample. The measured concentration of the
spiked sample in the assay was compared to the expected concentration. The average recovery was 91%.
E. Specificity
Percent of cross reactivity
Human insulin: 100%
Rat insulin: 100%
Publications Citing This Product
1. Jiang X, Zhou Y, Wu KK, Chen Z, Xu A, Cheng KK. APPL1 prevents pancreatic beta cell death and inflammation by dampening
NFκB activation in a mouse model of type 1 diabetes. Diabetologia. 2017 Mar 1;60(3):464-74.
2. Geng L, Liao B, Jin L, Huang Z, Triggle CR, Ding H, Zhang J, Huang Y, Lin Z, Xu A. Exercise alleviates obesity-induced
metabolic dysfunction via enhancing FGF21 sensitivity in adipose tissues. Cell reports. 2019 Mar 5;26(10):2738-52.
3. Yang BC, Wu SY, Leung PS. Alcohol ingestion induces pancreatic islet dysfunction and apoptosis via mediation of FGF21
resistance. Annals of translational medicine. 2020 Mar;8(6).
4. Sun H, Wei G, Liu H, Xiao D, Huang J, Lu J, Miao J, Liu J, Chen S. Inhibition of XBP1s ubiquitination enhances its protein stability and improves glucose homeostasis. Metabolism. 2020 Apr 1;105:154046.
5. Breznik JA, Foley KP, Maddiboina D, Schertzer JD, Sloboda DM, Bowdish DM. Effects of Obesity-Associated Chronic
Inflammation on Peripheral Blood Immunophenotype Are Not Mediated by TNF in Female C57BL/6J Mice. ImmunoHorizons. 2021
Jun 4;5(6):370-83.