Applera Corporation

Tropix, Inc.

Life Technologies Corporation

Anthony C Chiulli

Irena Y Bronstein

John C Voyta

Michelle A J Palmer

A competitive assay to determine the presence and concentration of an intracellular analyte (e.g., cAMP) in a sample is provided. All of the steps of the assay can be performed on the same assay plate, thereby eliminating the need to transfer the cells from a tissue culture plate on which the cells are grown, induced and lysed to a separate assay plate. The assay procedure includes combining, in a reaction chamber provided with a capture antibody, an antibody for the analyte, the sample to be assayed, and a conjugate of the analyte and an enzyme such as alkaline phosphatase. The mixture is incubated and washed and an enzyme labile substrate (e.g., a chemiluminescent, fluorescent or calorimetric substrate) is added. The assay can also be performed with a tagged analyte (e.g., an analyte having a radioactive or fluorescent tag) instead of an enzyme conjugate.

Cell growth, induction and lysis in an antibody-coated microplate for use in an ELISA

A competitive assay for the concentration of a cyclic nucleotide such as cyclic adenosine monophosphate combines, in a reaction chamber provided with a capture antibody, an antibody for the cAMP or other cyclic nucleotide, the sample to be assayed and a conjugate of cAMP and alkaline phosphatase. The mixture is incubated and washed, and after washing, a 1,2-dioxetane which is caused to decompose when contacted by the alkaline phosphatase of said conjugate is added. The light emission caused by decomposition is measured, with the strength of the signal being inversely related to the concentration of the cyclic nucleotide. Optionally, an enhancement agent in the form of a polymeric onium salt may be added, to further enhance the light emission.

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Maynard, MA, United States of America

Anthony C Chiulli