Development of a multipurpose time-resolved fluorescence immunoassay for rat insulin

Farah Van Genderen, Frans Gorus, Ilse Vermeulen, Evilien Vekens, Pieter DE PAUW, Daniel Pipeleers, Christiaan Van Schravendijk

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

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 25mul could be analyzed in less than 2days with a measuring range between 5 and 1250pg (0.2-50mug/L or 34.4-8600pM). The inter- and intraassay percentage coefficients of variation were less than 8.3 and 5.1, respectively. Recoveries of 0.48 to 40mug/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
Original languageEnglish
Pages (from-to)8-13
Number of pages6
JournalAnal Biochem
Volume404
Publication statusPublished - 2010

Keywords

  • rat insulin
  • immunoassay

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