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Luc-Pair™ Duo-Luciferase HT Assay Kit

For High-Throughput Applications

Introduction

GeneCopoeia's Luc-Pair™ Duo-Luciferase HT Assay Kit is a convenient system for high-throughput detection of Firefly luciferase (FLuc) and Renilla luciferase (Rluc) activities in 96- or 384-well plates. The Luc-Pair™ Duo-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 0–10% serum (e.g. DMEM, RPMI1640, etc.) Firefly luciferase luminescence is produced by one reagent, while a second reagent simultaneously quenches the Firefly luciferase and produces Renilla luciferase luminescence. 

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

 

Advantages

· Enhanced stability. 
· Convenient — Directly lyse cells in culture medium and measure luciferase activities simultaneously. 
· Versatility — Works well with many different eukaryotic (adherent or suspended) cells using micro-plate luminescence readers.
· Low background — Very limited background luminescence. No subtraction from readings is required.
· Simplicity — Renilla luciferase buffer contains the quenchers for Firefly luciferase activity (Figure 3). enabling a quick Glow and Stop-N-Glow two-step assays.
· Reproducibility — Reliable, linear results for a concentration range over several orders of magnitude. 

 

Figure 1.  Activities of Firefly and Renilla luciferase signals using the GeneCopoeia (GCI) Luc-Pair Duo-Luciferase HT Assay Kit. HEK 293 cells were transfected with Promega pGL4.13/pGL4.75 reporter vectors for 48 hours. FLuc and RLuc activities were measured as described in the procedure. Promega’s Dual-Glo luciferase assay kit was used (FLuc-Prmg and RLuc-Prmg) in comparison.  

 

Figure 2. Firefly and Renilla luciferase activity mesured in various media. HEK 293 cells were transfected with Promega pGL4.13/pGL4.75 reporter vectors for 48 hours. 5×104 transfected cells were suspended in DMEM without serum or the following types of media containing 10% serum:  DMEM, RPMI1640, EMEM, IMDM, McCoy’s 5A, F-12K, MEBM, ACL-4, L15, Cho-S-SFMII, NCTC109. Firefly and Renilla luciferase activities were measured as described in the procedure.

Figure 3. Firefly luciferase activity is quenched with the addition of Luc-HT Buffer II. HEK 293 cells were transfected with Promega pGL4.13/pGL4.75 reporter vectors for 48 hours. Firefly luciferase activity was measured as described in the procedure. Next, 1× Luc-HT 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

Luc-Pair™ Duo-Luciferase HT Assay Kit

Buy Catalog# Product Description Price
LF013 Luc-Pair™ Duo-Luciferase HT Assay Kit (10 ml) For high-Throughput detection of Firefly and Renilla luciferase activities. $156
LF014 Luc-Pair™ Duo-Luciferase HT Assay Kit (30 ml) $375
LF015 Luc-Pair™ Duo-Luciferase HT Assay Kit (100 ml) $1022

  User manual MSDS

 

Vector options (Discounted price – $199)

Buy Catalog# Product Description Promoter Reporter gene Tracking gene Selection marker Size Price
ZX001 pEZX-FR01 Firefly luciferase (FLuc) and Renilla luciferase (Rluc) reporter cloning vector N/A* Firefly luciferase (hLuc) Renilla luciferase (Rluc) Puromycin 10µg $495
ZX002 pEZX-FR02 Firefly luciferase (FLuc) and Renilla luciferase (Rluc) reporter cloning vector SV40 Firefly luciferase (hLuc) Renilla luciferase (Rluc) N/A* 10µg $495
ZX003 pEZX-FR03 Firefly luciferase (FLuc) and Renilla luciferase (Rluc) reporter cloning vector miniCMV Firefly luciferase (hLuc) Renilla luciferase (Rluc) Puromycin 10µg $495

 

 

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.