Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase
1996; Future Science Ltd; Volume: 21; Issue: 3 Linguagem: Inglês
10.2144/96213pf01
ISSN1940-9818
AutoresChris Martin, P.A.L. Wight, Anna Dobretsova, Irena Bronstein,
Tópico(s)Photoreceptor and optogenetics research
ResumoA unique combined luminescence assay for firefly (Photinus pyralis) luciferase and β-galactosidase (β-gal) reporter gene products is described. Luciferase and β-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-LightTM assay system, luciferase activity is measured first with an enhanced luciferase assay, followed by quantitation of β-gal with Galacton-PlusTM chemiluminescent substrate and Sapphire-IITM enhancer. Highly sensitive detection of luciferase (2 fg) and β-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 β-gal reporter genes is also described.
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