Choosing a Luciferase Reporter Assay

When creating a luciferase reporter assay, there are two important elements to consider: the luciferase reporter protein itself and the assay chemistry used to detect reporter activity. The characteristics of these two components together contribute to the overall performance of the assay. By selecting the reporter/detection solution that is optimal for your experimental goals, you can customize your luciferase reporter assay to create the best solution for your research.

Promega offers a choice of 3 different luciferase reporters: NanoLuc® Luciferase (Nluc,19kDa), Renilla Luciferase (Rluc, 36kDa) and Firefly Luciferase (Fluc, 61kDa), which vary in size, brightness and protein half-life.

Multiple assay detection reagents are also available for each reporter. Key considerations for selecting the optimal assay reagent include:

• Signal intensity and overall dynamic range needed for the assay.
• Signal stability, or half-life, which will impact your processing workflow.
• Processing steps required. Non-homogenous assays require a separate lysate creation step prior to reagent addition. Homogenous assay reagents are added directly to the cells in culture eliminating sample pre-processing.
• Lytic or live-cell reporter detection
• Single or dual-reporter detection