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Methods Enzymol ; 326: 175-202, 2000.
Article in English | MEDLINE | ID: mdl-11036643

ABSTRACT

Chemiluminescent reporter gene assays provide highly sensitive, quantitative detection in simple, rapid assay formats for detection of reporter enzymes that are widely employed in gene expression studies. Chemiluminescent detection methodologies typically provide up to 100-1000x higher sensitivities than may be achieved with fluorescent or colorimetric enzyme substrates. The variety of chemiluminescent 1,2-dioxetane substrates available enable assay versatility, allowing optimization of assay formats with the available instrumentation, and are ideal for use in gene expression assays performed in both biomedical and pharmaceutical research. In addition, 1,2,-dioxetane chemistries can be multiplexed with luciferase detection reagents for dual detection of multiple enzymes in a single sample. These assays are amenable to automation with a broad range of instrumentation for high throughput compound screening.


Subject(s)
Alkaline Phosphatase/analysis , Genes, Reporter , Glucuronidase/analysis , Luciferases/analysis , beta-Galactosidase/analysis , 3T3 Cells , Alkaline Phosphatase/genetics , Animals , Cell Adhesion , Cell Culture Techniques/methods , Cells, Cultured , Glucuronidase/genetics , Humans , Indicators and Reagents , Luciferases/genetics , Luminescent Measurements , Mammals , Mice , Recombinant Fusion Proteins/analysis , Recombinant Fusion Proteins/genetics , Transfection , beta-Galactosidase/genetics
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