Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase | Zendy

Chris Martin | Zendy; P.A.L. Wight | Zendy; Anna Dobretsova | Zendy; Irena Bronstein | Zendy

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Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase

Author(s) -

Chris Martin,

P.A.L. Wight,

Anna Dobretsova,

Irena Bronstein

Publication year - 1996

Publication title -

biotechniques

Language(s) - Uncategorized

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/96213pf01

Subject(s) - luciferase , reporter gene , luciferases , microbiology and biotechnology , luciferin , bioluminescence , enhancer , chemistry , bioreporter , chemiluminescence , biology , gene , gene expression , biochemistry , transfection , chromatography

A unique combined luminescence assay for firefly (Photinus pyralis) luciferase and beta-galactosidase (beta-gal) reporter gene products is described. Luciferase and beta-gal activities are determined with the same aliquot of cell lysate prepared from cells contransfected with both reporter genes, thereby reducing manual labor and increasing experimental accuracy. With the Dual-Light assay system, luciferase activity is measured first with an enhanced luciferase assay, followed by quantitation of beta-gal with Galacton-Plus chemiluminescent substrate and Sapphire-II enhancer. Highly sensitive detection of luciferase (2 fg) and beta-gal (8 fg) is achieved with a dynamic range over seven orders of magnitude of enzyme concentration. Comparative analysis of both independent and combined (Dual-Light) detection methods for cells contransfected with luciferase and beta-gal reporter genes is also described.

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