The Dual-Luciferase® Reporter (DLR™) Assay System provides an efficient means of performing dual reporter assays. In the DLR™ Assay, the activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis, also known as sea pansy)luciferases are measured sequentially from a single sample. The firefly luciferase reporter is measured first by adding Luciferase Assay Reagent II (LAR II) to generate a “glow-type” luminescent signal. After quantifying the firefly luminescence, this reaction is quenched, and the Renilla luciferase reaction is initiated by simultaneously adding Stop & Glo® Reagent to the same tube. The Stop & Glo® Reagent also produces a “glow-type” signal from the Renilla luciferase, which decays slowly over the course of the measurement. In the DLR™ Assay System, both reporters yield linear assays with subattomole (<10-18)sensitivities and no endogenous activity of either reporter in the experimental host cells. Furthermore, the integrated format of the DLR™ Assay provides rapid quantitation of both reporters either in transfected cells or in cell-free transcription/translation reactions.
Promega has two series of firefly and Renilla luciferase vectors, pGL4 and pRL, designed for use with the DLR™ Assay Systems. These vectors may be used to co-transfect mammalian cells with any experimental and control reporter genes
.
Dual-Luciferase® Reporter (DLR™) Assay System (Dual-Luciferase® 双萤光素酶报告基因检测系统) 为双报告基因检测提供有效的手段。 在DLR™ 检测中，萤火虫(Photinus pyralis ) 萤光素酶和海肾(Renilla reniformis ) 萤光素酶的活性可在单个样品中依次检测。首先检测萤火虫萤光素酶，将萤光素酶检测试剂II(Luciferase Assay Reagent II, LAR II)加入样品中，产生的光信号至少持续1分钟。定量萤火虫萤光强度后，再在同一个样品中加入Stop & Glo® 试剂，将上述反应终止，同时启动海肾萤光素酶反应。使用带有试剂自动注射器的发光检测仪，可在4 秒内完成两个检测。在DLR™ 检测系统中，两个报告基因产生的线性检测的灵敏度均可达埃摩尔级(<10-18)，在实验宿主细胞内均无内源活性。而且，DLR™ 检测的一体化模式既可快速定量检测转染细胞，也可用于快速定量检测无细胞转录/ 翻译反应体系中的两个报告基因。
为了得到Dual-Luciferase® 试剂盒的最佳检测结果，Promega推荐使用针对该检测过程进行过确证的发光检测仪。这些发光检测仪均带有DLReady ™ 标识。请参阅“通过DLReady ™ 确证的发光检测仪”页面，来了解相应的仪器信息。
pGL4 萤光素酶报告基因载体是针对DLR™ 检测系统而设计。组成型表达的海肾萤光素酶载体可与任何实验用萤火虫萤光素酶载体联合共转染哺乳动物细胞。
Cat.# E1960 和E1980 产品请注意：系统中已提供了足够的Passive Lysis Buffer，能够进行96 孔板细胞的1,000 次检测(一般每孔用
20μl 1X PLB)。若需要更多的Passive Lysis Buffer( 如>100μl/ 孔)，可单独购买。