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Free and total anti-insulin antibody radioimmunoassay kit

Free and total anti-insulin antibody radioimmunoassay kit RIA

The AAI kit is a RIA assay to measure the free and total anti-insulin autoantibodies in human serum and plasma with regard to diabetes and hypoglycemia.

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The AAI kit is a RIA assay to measure the free and total anti-insulin autoantibodies in human serum and plasma with regard to diabetes and hypoglycemia.

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Overview

Predict the future development of type I diabetes in asymptomatic children, adolescents, and young adults

Differentially diagnose between of type I and type II diabetes Evaluate insulin resistance in patients with established diabetes (type I or type II)

Investigate hypoglycemia in non-diabetic subjects

Detect anti-insulin antibodies induced during a treatment by human or porcine insulin

Benefits

ACCURATE AND RELIABLE ASSAY

Assay principle: sandwich

Anti-insulin antibody (AIAb) detection is based on the demonstration of a specific binding with iodine-125 radiolabeled insulin.

There are two alternative protocols to measure levels of free AIAb or total AIAb.

1- Free anti-insulin antibodies The free AIAb are antibodies not complexed with circulating insulin. They are determined by a radioimmunoprecipitation assay in liquid phase. 50µL of serum containing anti-insulin antibodies are incubated with 125I-radiolabeled insulin. The 125I-insulin bound to the AIAb fraction is separated from the free 125I-insulin fraction by precipitation with PEG. After centrifuging, radioactivity counting of the pellets (bound fraction) enables the calculation of the binding percentage of 125I-insulin.

Assay procedure: 2 h at RT + 10 min at RT + 10 min centrifugation

2- Total anti-insulin antibodies The total AIAb are the free AIAb plus the antibodies complexed with serum insulin. They are determined by following the same protocol as the free AIAb after dissociation of immune complexes in acid medium, insulin adsorption by active charcoal, neutralization and then centrifuging. 1- Extraction by active charcoal 200µl sample Procedure: 10 min at RT + 10 min centrifugation 2- Immunoprecipitation 100µl supernatant Assay procedure: 2 h at RT + 10 min at RT + 10 min centrifugation