Dual-Luciferase? Reporter Assay System

产品编号：E1910 品牌：promega 原厂货号：E1910
产品分类：蛋白质检测与分析 > 酶检测和活性分析 > 荧光素酶检测
应用分类：蛋白质研究 > 报告基因检测 > 荧光素酶报告基因检测

包装：

100 assays

运保温度:

储存于-20℃。

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其它组分：

Luciferase Assay Substrate【子货号：E151A，包装：1 x 1 vial，，运保温度：-20℃】

Luciferase Assay Buffer II【子货号：E195A，包装：1 x 10ml，，运保温度：-20℃】

Stop & Glo® Substrate【子货号：E640A，包装：1 x 200μl，，运保温度：-20℃】

Stop & Glo® Buffer【子货号：E641A，包装：1 x 10ml，，运保温度：-20℃】

Passive Lysis Buffer, 5X【子货号：E194A，包装：1 x 30ml，，运保温度：-20℃】

描述：

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。

原厂资料：

The Dual-Luciferase® Reporter (DLR™) Assay System provides an efficient means of performing two reporter assays. In the DLR™ Assay, the activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis or 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 luminescent signal lasting at least one minute. After quantifying the firefly luminescence, this reaction is quenched, and the Renilla luciferase reaction is initiated simultaneously by adding Stop & Glo® Reagent to the same sample. Both assays can be completed in about 4 seconds using a luminometer with reagent auto-injectors. In the DLR™ Assay System, both reporters yield linear assays with attomole (<10^–18) sensitivities and no endogenous activity 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.

For best results with the Dual-Luciferase® Assay, Promega recommends using a luminometer that has been validated for use with the assay. These luminometers are qualified as DLReady™. For a listing of qualified instruments, please visit the DLReady™ Validated Luminometers page.

The pGL4 Luciferase Reporter Vectors are designed for use with the DLR™ Assay Systems. A Renilla luciferase vector with constitutive expression may be used in combination with any experimental firefly luciferase vector to co-transfect mammalian cells.

Notice for Cat.# E1960 and E1980: Sufficient Passive Lysis Buffer is provided to perform 1,000 assays with cells grown in 96-well plates (typically 20μl of 1X PLB per well). For applications requiring more lysis reagent (e.g., >100μl/well), additional Passive Lysis Buffer may be purchased separately.

Features - Benefits

Greater Accuracy: Renilla luciferase internal control allows more accurate results.
Convenience: Samples don't have to be split; saves plates and time.
Sensitivity: Allows study of weak promoters, low-level expression/regulation and expression in cells that transfect poorly.
Linearity: Range extends 7 logs; very active samples typically do not need dilution.

Applications

For more information, see the Protocols & Applications Guide.

注意事项：

储存条件

储存于-20℃。