High-sensitive Plus Mouse Insulin ELISA Kit

Cat No.: 32393          ★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 co-incubated 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.

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<10%.

Inter-assay Precision (Precision between assays) C.V<10%.

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%.

D. Specificity

Percent of cross reactivity

Human insulin: 100%

Rat insulin: 100%

E. Validation

Serum insulin levels were measured in two group of 6 hours fasting C57 mice. (HFD-fed & STC-fed)