Dual-Luciferase Reporter Assay System 10-Pack

Description

• Overview
• Protocols
• Specifications
• Resources

• Sensitive, Rapid and Convenient Dual-Reporter Assay

  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, we recommend 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 20ul of 1X PLB per well). For applications requiring more lysis reagent (e.g., >100ul/well), additional Passive Lysis Buffer may be purchased separately.

  Assay Advantages

  • Normalization to Renilla luciferase internal control allows more accurate results.
  • Samples don't have to be split, saving plates and time.
  • Allows study of weak promoters, low-level expression/regulation and expression in cells that transfect poorly.
  • Linear over 7 logs; very active samples typically do not need dilution.

• Protocols

  Complete Protocol

  

  Dual-Luciferase® Reporter Assay System Technical Manual

  PDF (641 KB)

  

  Dual-Luciferase® Reporter 1000 Assay System Technical Manual

  PDF (514 KB)

  Quick Protocols

  

  Dual-Luciferase Reporter Assay and Dual-Luciferase Reporter 1000 Assay System Quick Protocol FB024

  PDF (73 KB)

  Video Protocols

  

• Certificate of Analysis

  Lookup Certificate of Analysis

  Storage Conditions

  -30C TO -10C

  Use Restrictions

  For Research Use Only. Not for Use in Diagnostic Procedures.

• Resources

  

  Articles

  • Designing a Bioluminescent Reporter Assay: Normalization
  • Measuring the Dual-Luciferase Reporter Assay on the GloMax® Discover System
  • Multiplexing Cell-Based Assays: Get More Biologically Relevant Data
  

  Citations

  • Loss of BRCA1 in the cells of origin of ovarian cancer induces glycolysis: A window of opportunity for ovarian cancer chemoprevention.
    --
    2017 Cancer Prev. Res.