AffinityImmuno Products

Introduction

Amylin, encoded by IAPP, is a 37-amino-acid peptide hormone of the calcitonin family that is co-secreted with insulin from pancreatic beta cells in response to meals. Acting mainly through calcitonin receptor complexes paired with receptor-activity-modifying proteins, it slows gastric emptying, suppresses inappropriate glucagon release after eating, promotes satiety, and selectively blunts insulin-stimulated glucose uptake in skeletal muscle without affecting fat cell metabolism. Circulating amylin is measurably diminished in type 1 diabetes and in advanced type 2 diabetes as beta-cell mass declines, yet within the pancreas itself the peptide has a tendency to misfold and accumulate as islet amyloid, a process thought to further accelerate beta-cell loss in type 2 disease. The hormone also has reported effects on bone remodeling and blood pressure regulation, along with an intrinsic antimicrobial activity, reflecting functions that extend beyond glucose control alone.

Amylin’s tendency to misfold and accumulate as islet amyloid is thought to accelerate beta-cell loss in type 2 diabetes, so this assay supports both diabetes biomarker studies tracking declining amylin secretion and islet-biology research into amyloid-driven beta-cell dysfunction.

Principle of the Assay

This kit utilizes a competitive inhibition enzyme immunoassay format to measure Human ProIAPP. Microtiter plate wells are pre-coated with Human ProIAPP antigen. During the assay, standards or samples are added simultaneously with a biotin-conjugated antibody specific to Human ProIAPP; free ProIAPP present in the standard or sample competes with the plate-bound antigen for a limited supply of this detection antibody. Streptavidin-conjugated Horseradish Peroxidase (HRP) is then added to each well and allowed to incubate, linking enzymatic activity to antibody captured on the plate. Following incubation, TMB substrate solution is added to initiate the colorimetric enzyme-substrate reaction. The reaction is stopped by addition of a sulfuric acid stop solution, and absorbance is measured spectrophotometrically at 450 nm (± 10 nm). Because free analyte in the sample competes with plate-bound antigen for antibody binding, the resulting signal is inversely proportional to ProIAPP concentration; sample concentrations are determined by comparing optical density values against the standard curve.