Luc-Pair Duo-Luciferase HT Assay Kit
| Specifications | |
|---|---|
| Product Category: | Dual Reporter Assays: Firefly Luciferase and Renilla Luciferase |
Product Description
- Enhanced stability
- Directly lyse cells in culture medium and measure luciferase activities simultaneously
- For many different eukaryotic (adherent or suspended) cells using micro-plate luminescence readers
- Very limited background luminescence. No subtraction from readings is required
- Quenched Firefly luciferase activity (Figure 2) for quick Glow and Stop-N-Glow two-step assays
- Reliable, linear results for a concentration range over several orders of magnitude
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.
Performance Data
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).
