luciferriseassay介绍.pdfVIP

Promega Notes Magazine Number 57, 1996, p.02 Dual-LuciferaseTM Reporter Assay: An Advanced Co-Reporter Technology Integrating Firefly and Renilla Luciferase Assays By Bruce A. Sherf, Shauna L. Navarro, Rita R. Hannah and Keith V. Wood Promega Corp oration In the quantitation of gene expression using firefly luciferase, a second reporter gene is commonly used to minimize experimental variability. However, traditional co-reporters (e.g., CAT, beta-Gal or GUS) are inconvenient due to differences in their respective assay chemistries, handling requirements and measurement characteristics. Promega introduces a superior dual-rep orter technology integrating the assay offirefly luciferase with the Renilla lucif erase assay . The Dual-LuciferaseTM Rep orter Assay System *, in combination with the pRL Vectors which express Renilla luciferase as the second reporter, offers the exceptional speed, sensitivity and convenience of two luciferase reporter assays in a single-tube format. The system also includes Passive Lysis Buffer, formulated to provide quantitative solubilization of both luciferases from mammalian cells cultured in multi -well plates without the need for TM individual processing of each sample. The Dual-Luciferase Reporter Assay System will provide immediate benefit to researchers currently using any exp erimental firef ly lucif erase rep orter vector. *Patent pending. Introduction Dual-reporters are used to make relational or ratiometric measurements within an experimental system. Typically one reporter is used as an internal control to which measurement of the other reporter is normalized. In measurements of gene expression, dual reporters are generally used in transient transfections of cultured cells, where one vector containing the experimental reporter gene is co-transfected with a second vector containing a distinct reporter gene serving as the control. Usual