Introduction
GeneCopoeia’s Luc-Pair™ Firefly Luciferase Assay Kit is the perfect choice for high sensitivity detection of Firefly luciferase (FLuc) activity. The Luc-Pair™ Firefly Luciferase HS Assay Kit works great for many different eukaryotic cell types (e.g. mammalian, invertebrates, etc.) using micro-plate or single-tube luminometers, and is compatible with GeneCopoeia, Promega and many other Fluc vectors.
Advantages
- High sensitivity. The reagents have been developed so that the signals for Firefly luciferase exhibit greatest sensitivity (Figure 1).
- Versatility. The system has been designed for assays with many different eukaryotic (vertebrates, lower invertebrates) cells using micro-plates or single-tube luminescence reader.
- Low background. The system produces very limited background luminescence. No subtraction is required from readings.
- Reproducibility. This system is designed to yield reliable, linear results for a concentration range over several orders of magnitude.
GeneCopoeia also offers the Luc-Pair™ Duo-Luciferase Assay Kit 2.0 (for enhanced stability) and Luc-Pair™ Duo-Luciferase HS Assay Kit (for high sensitivity) for applications requiring luciferase readings.
Figure 1. Activity of Firefly luciferase signals using GeneCopoeia Luc-Pair™ Firefly Luciferase HS Assay Kit. HEK 293 cells were transfected with Promega pGL4.13 reporter vector for 48 hours. The FLuc (FLuc-GeneCopoeia) activity was measured as described in the procedure. Promega’s Luciferase Assay System was used (FLuc-Promega) in comparison.
Figure 2. Signal stability of luciferase using GeneCopoeia Luc-Pair Firefly-Luciferase (FLuc) HS Assay Kit. HEK 293 cells were transfected with Promega pGL4.13 reporter vector for 48 hours. The FLuc (FLuc-GeneCopoeia) activity was measured as described in the procedure. A sample injector was used to dispense the Fluc Working Solution, set shaking to mix for 1 sec, and counting was started immediately for 20 sec.
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.
