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Free & total anti-insulin antibody radioimmunoassay kit

Free & total anti-insulin antibody radioimmunoassay kit RIA

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

For Research Use Only, not for use in diagnostic procedures.

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

For Research Use Only, not for use in diagnostic procedures.

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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