Cisbio Bioassays offers an accurate enzymatic (RIA) assay for measuring the free and total anti-insulin autoantibodies. It is based on the sandwich type principle and carried out in human serum and plasma.
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Free and total anti-insulin antibody (AIAb) radioimmunoassay kit
Features
- Working range: negative & positive controls
- Sample: serum, plasma
Applications
- Predicting the future development of type 1 diabetes in asymptomatic children, adolescents, and young adults
- Differential diagnosis of type 1 versus type 2 diabetes
- Evaluating diabetics with insulin resistance in patients with established diabetes (type 1 or type 2)
- Investigation of hypoglycemia in non-diabetic subjects
- Detection of anti-insulin antibodies induced during a treatment by human or porcine insulin
Assay principle
RIP
Immunoprecipitating reagent
The principle of anti-insulin antibody (AIAb) detection is based on the demonstration of a specific binding with iodine- 125 radiolabeled insulin. Two alternative protocols.
1-Free anti-insulin antibodies (Free AIAb)
The free AIAb are antibodies not complexed with circulating insulin. They are determined by a radioimmunoprecipitation assay in liquid phase: - 50?L serum containing anti-insulin antibodies is incubated with 125I-radiolabeled insulin. - the 125I-insulin bound to the AIAb fraction is separated from the free 125I-insulin fraction by precipitation with polyethylene glycol (PEG). - after centrifuging, radioactivity counting of the pellets (bound fraction) enables the calculation of the binding percentage of 125I-insulin.
*Three-step assay procedure: 2 h at RT + 10 min at RT + 10 min centrifugation.
2- Total anti-insulin antibodies (Total AIAb)
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, and then centrifuging.
21- Extraction by active charcoal ? 200?l sample Assay procedure: 10 min at RT + 10 min centrifugation
22- Immunoprecipitation 100?l supernatant Assay procedure: 2 h at RT + 10 min at RT + 10 min centrifugation.