Mouse CYSTM1 Immunoassay Kit
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 quantitative sandwich ELISA. The microplate is pre-coated with a polyclonal antibody specific for mouse CYSTM1. Standards and samples are pipetted into the wells and any mouse CYSTM1 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-labelled polyclonal antibody specific for mouse CYSTM1 is added to the wells. After wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP-substrate solution is added and colour develops in proportion to the amount of mouse CYSTM1 bound initially. The assay is stopped and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured mouse CYSTM1, the unknown sample concentration can be interpolated from a reference curve included in each assay.
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each assay.
The lowest insulin level that can be measured by this assay is 0.025 ng/ml.
Intra-assay Precision (Precision within an assay) C.V. < 10%.
Inter-assay Precision (Precision between assays) C.V. <10%.
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 92%.
Percent of cross reactivity
Human insulin 100%
Rat insulin 100%
Insulin levels were measured at various time points after intra-peritoneal glucose challenge in overnight fasting C57 mice (standard chow fed).