Patent ID: 6686171
Filing Date: 2004-02-03
Classification: G01N

Abstract:
A competitive assay for determining the concentration of a cyclic nucleotide phosphate in a sample comprising cells, the assay comprising:lysing cells in the sample; combining, in a reaction chamber provided with a capture antibody, (1) a conjugate of said cyclic nucleotide phosphate and an alkaline phosphatase, (2) a primary antibody that is bound by said capture antibody and that binds, when so bound, said cyclic nucleotide phosphate, and (3) said sample to form a reaction mixture and incubating the reaction mixture so as to permit binding of said primary antibody and said conjugate, washing said reaction mixture to remove any unbound conjugate or antibody, adding to said reaction mixture an alkaline phosphatase-triggered 1,2-dioxetane which, when contacted with alkaline phosphatase in said conjugate, is caused to decompose and release light, and detecting any chemiluminescent signal emitted by said reaction mixture, wherein said signal is indicative of the concentration of cyclic nucleotide phosphate in the sample.