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Luc-Pair™ Firefly Luciferase HT Assay Kit

– For High-Throughput Applications

Introduction

GeneCopoeia's Luc-Pair™ Firefly Luciferase HT Assay Kit is a convenient system for high-throughput detection of Firefly luciferase (FLuc) activity in 96- or 384-well plates. The Luc-Pair™ Firefly Luciferase HT Assay reagents can be added directly to cells in growth medium without washing or preconditioning, and are optimized for use with many different media containing serum (e.g. DMEM, RPMI1640, etc.).

The Luc-Pair™ Firefly Luciferase HT Assay Kit works great for many different eukaryotic cell types (e.g. mammalian, invertebrates, etc.)  and is compatible with many Fluc vectors from GeneCopoeia, Promega and other sources.

 

Advantages

· Enhanced stability – Greater signal stability, with a half-life of approximately 2 hours (Figure 1). Ideal  for high throughput assays.
· Convenience — Directly lyse cells in culture medium and measure luciferase activity simultaneously.
· Flexibility — Works well in many types of culture media, even with serum present (Figure 2).
· Versatility — Works well with many different eukaryotic (adherent or suspension) cells using micro-plate luminescence readers.
· Low background — Very limited background luminescence. No subtraction from readings is required.
· Reproducibility — Reliable, linear results for a concentration range over several orders of magnitude.

 

The Luc-Pair™ Firefly Luciferase HT Assay Kit provices excellent FLuc signal stability, with a half life of approximately 2 hours (Figure 1).

 

Figure 1.  Stability of Firefly luciferase signal. HEK 293 cells were transfected with Promega pGL4.13 reporter vector for 48 hours. Fluc activity was measured at various time points using the Luc-Pair FLuc HT Assay Kit as described in the procedure. 

 

The Luc-Pair™ Firefly Luciferase HT Assay Kit allows excellent signal strength and stability in many different types of culture media, even with serum present (Figure 2).

 

Figure 2. Firefly luciferase activities in various media. HEK 293 cells were transfected with Promega pGL4.13/pGL4.75 reporter vectors for 48 hours. 5×104 transfectd cells were suspended in DMEM without serum or the following types of media containing 10% serum:  DMEM, RPMI1640, EMEM, IMDM, McCoy’s5A, F-12K, MEBM, ACL-4, L15, Cho-S-SFMII, NCTC109. Firefly luciferase activities were measuredat various time points using the Luc-Pair FLuc HT Assay Kit as described in the procedure.