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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.

 

 

To Order

Luc-Pair™ Firefly Luciferase HT Assay Kit

Buy Catalog# Product Description Price
LF016 Luc-Pair™ Firefly Luciferase HT Assay Kit (10 ml) For high-Throughput detection of Firefly luciferase activities. $80
LF017 Luc-Pair™ Firefly Luciferase HT Assay Kit (30 ml) $130
LF018 Luc-Pair™ Firefly Luciferase HT Assay Kit (100 ml) $390

  User manual MSDS
 

FAQs

Frequently Asked Questions: Luciferase Assays

Answer: “RLU” stands for “Relative Light Unit”. Luminometers do not measure light output, in terms of numbers of photons, directly.
Answer: All GeneCopoeia luciferase kits are compatible with all known luminometers.
Answer: All GeneCopoeia luciferase reagents are stable for at least 6 months if stored at -20°C.
Answer: To prevent loss of activity, it is best to avoid repeated freezing and thawing. We recommend pipetting small aliquots into new, clean, sterile tubes and placing those aliquots at -20°C. Once you have thawed one of the aliquots, discard it instead of putting it back in the freezer.
Answer: The Gaussia luciferase substrate, coelenterazine, is also the substrate for Renilla luciferase, so it is compatible with Renilla. However, coelenterazine cannot be used with Firefly (Photinus pyralis) luciferase, which instead uses luciferin as its substrate.
Answer: Unfortunately, the individual components of GeneCopoeia's luciferase kits cannot be purchased separately at this time.
Answer: Typically, between 12 and 24 hours.
Answer: The Luc-Pair™ Duo-Luciferase Assay Kit 2.0 (For Enhanced Stability), Luc-Pair™ Duo-Luciferase Assay Kit HT (For High Throughput), Luc-Pair™ Firefly Luciferase HT Assay Kit, and Luc-Pair™ Renilla Luciferase HT Assay Kit (For High Throuhput), which provide greater signal stability, are most useful when assaying a relatively large number of samples. Their enhanced signal stability means that there will be little sample-to-sample variation due to signal decay. Conversely, the Luc-Pair™ Duo-Luciferase HS Assay Kit (For High Sensitivity), Luc-Pair™ Firefly Luciferase HS Assay Kit (For High Sensitivity), and Luc-Pair™ Renilla Luciferase HS Assay Kit (For High Sensitivity) are used when maximal signal intensity is required, but the trade-off is that the signal decays more rapidly under typical laboratory conditions. These kits are not appropriate for measuring the activities of large numbers of samples, unless sample injectors are used.