Development of a multipurpose time-resolved fluorescence immunoassay for rat insulin
| Autor: | Frans Gorus, Evilien Vekens, Farah T. van Genderen, Pieter De Pauw, Ilse Vermeulen, Daniel Pipeleers, Chris Van Schravendijk |
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| Přispěvatelé: | Diabetes Pathology & Therapy, Medical Biochemistry, Pathologic Biochemistry and Physiology, Pathology/molecular and cellular medicine |
| Předmět: |
Streptavidin
Time Factors medicine.drug_class medicine.medical_treatment Fluoroimmunoassay Biophysics Monoclonal antibody Biochemistry Microtiter plate chemistry.chemical_compound Europium medicine Animals Insulin Rats Wistar Molecular Biology Detection limit Immunoassay Chromatography medicine.diagnostic_test Chemistry Antibodies Monoclonal Radioimmunoassay Cell Biology Molecular biology Rats Biotinylation rat insulin |
| Zdroj: | Vrije Universiteit Brussel |
| Popis: | We present a time-resolved fluorescence immunoassay (TR-FIA) for the measurement of rat insulin in cell extracts and culture media. This assay is based on the binding of two monoclonal antibodies to different parts of the insulin molecule in a 96-well microtiter plate. For the detection, europium-labeled streptavidin that interacts with the second biotinylated antibody is used. Samples of 25 microl could be analyzed in less than 2 days with a measuring range between 5 and 1250 pg (0.2-50 microg/L or 34.4-8600 pM). The inter- and intraassay percentage coefficients of variation were less than 8.3 and 5.1, respectively. Recoveries of 0.48 to 40 microg/L rat insulin, added to culture medium, ranged between 94 and 107%. Results were significantly correlated with those of an in-house radioimmunoassay (RIA) for rodent insulin (P |
| Databáze: | OpenAIRE |
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