Introduction
GeneCopoeia's Luc-Pair™ Duo-Luciferase HS Assay Kit is the perfect choice for high sensitivity detection of Firefly luciferase (FLuc) and Renilla luciferase (Rluc) activities. The Luc-Pair™ Duo-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 and Rluc vectors. The Luc-Pair™ Duo-Luciferase HS Assay Kit is ideal for many applications, such as measuring the inhibitory effect of a miRNA on a target sequence.
Advantages
- High sensitivity — Reagents are optimized for greatest sensitivity of Firefly and Renilla luciferase signals (Figure 1).
- Versatility — Designed for assays with many different eukaryotic cell types (e.g. mammalian, lower invertebrates, etc.) using micro-plates or single-tube luminometers.
- Low background — Very low background luminescence. No subtraction is required from readings.
- Simplicity — Renilla luciferase buffer and substrate contains the quencher for Firefly luciferase activity (Figure 2), enabling quick Glow and Stop-N-Glow two-step assays.
- Reproducibility — Reliable, linear results for a concentration range over several orders of magnitude.
- Compatibility — Compatible with Promega's Fluc and Rluc vectors.
GeneCopoeia also offers the Luc-Pair™ Duo-Luciferase Assay Kit 2.0 for applications requiring luciferase readings with enhanced stability.
Figure 1. Activity of Firefly luciferase and Renilla luciferase signals using GeneCopoeia Luc-Pair™ Duo-Luciferase HS Assay Kit. HEK 293 cells were transfected with Promega pGL4.13/pGL4.75 reporter vectors for 48 hours. The FLuc (FLuc-GeneCopoeia) and RLuc (RLuc-GeneCopoeia) activity was measured as described in the procedure. The Promega dual-luciferase assay kit was used (FLuc-Promega and RLuc-Promega) for comparison.
Figure 2. Firefly luciferase activity is quenched with the addition of Luc-H Buffer II. HEK 293 cells were transfected with Promega pGL4.13/pGL4.75 reporter vectors for 48 hours. The firefly luciferase activity was measured as described in the procedure. Next, 1x Luc-H Buffer II (without or with substrate) was added to the wells, followed by count reading in a luminometer. About 99.9% of firefly luciferase activity was quenched (middle column).
To Order
Vector options (Discounted price – $199)
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.
Publications
View selected publications citing GeneCopoeia's Luc‐Pair™ Firefly/Renilla luciferase HS assay kit from the recent literature
2022
- Zheng, T., et al. (2022). MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma. Journal of Translational Medicine doi: 10.1186/s12967-021-03226- [Luc‐Pair™ Duo‐Luciferase HS Assay Kit]
- Tao, D., et al. (2022). CircPAG1 interacts with miR-211-5p to promote the E2F3 expression and inhibit the high glucose-induced cell apoptosis and oxidative stress in diabetic cataract. Cell Cycle doi: 10.1080/15384101.2021.2018213 – [Luc‐Pair™ Duo‐Luciferase HS Assay Kit]
- Zhang, D., et al. (2022). Circular RNA circRNF169 functions as a miR-30c-5p sponge to promote cellular senescence. Biochemical and Biophysical Research Communications doi: 10.1016/j.bbrc.2022.03.041 [Luc‐Pair™ Duo‐Luciferase HS Assay Kit]
- Kim, S.M., et al. (2022). Hyaluronan synthase 2, a target of miR-200c, promotes carbon tetrachloride-induced acute and chronic liver inflammation via regulation of CCL3 and CCL4. Experimental and Molecular Medicine doi: 10.1038/s12276-022-00781-5 [Luc‐Pair™ Duo‐Luciferase HS Assay Kit]
2021
