Application Notes & Technical Papers

Developing a Luciferase-Based NF-κB Reporter Assay with Transiently Transfected HEK 293 Cells

Firefly luciferase is widely used as a reporter mole-cule in cell based assays because of its sensitivity, dynamic range and ease of detection. Luciferase based assays have been developed for many dif-ferent applications, and they can be incorporated into multiple assay formats.

One of the challenges to developing a luciferase assay for high throughput drug screening involves the need to generate large numbers of cells that contain the reporter DNA. The most common ap-proach to cell based assay development involves creating stable cell lines, a costly, time consuming and labor intensive process that requires multiple rounds of selection and clonal isolation. In addi-tion, if an assay requires coexpression of reporter and target molecules, cells need to undergo addi-tional rounds of screening and exposure to multi-ple, harsh selection agents.

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