The Thermo Scientific Pierce Renilla Luciferase Flash Assay Kit provides researchers with a highly sensitive intracellular assay for transcriptional activity of regulatory elements in mammalian whole cell lysate.
This Flash Assay Kit contains reagents for measuring the activity of Renilla luciferase in mammalian cell lysates. When used with Thermo Scientific Green Renilla Luc Plasmids, the kit provides an extremely sensitive bioluminescent reporter assay system for intracellular detection of promoter or pathway activity. The signal produced by Green Renilla Luc is considerably greater than signal from either native firefly or Renilla luciferases assayed under similar conditions. Light output captured using a luminometer can be correlated with the amount of Renilla luciferase protein produced and used to determine the activity of the promoter driving Renilla expression.
- Sensitive – greater sensitivity allows utilization of smaller numbers of cells
- Cost effective – highest sensitivity assays result in decreased reagent consumption
- Time-saving – assays require minimal sample handling
- Automation-friendly – amenable to high throughput screens
- Convenient – contains a universal cell lysis buffer and optimized flash assay reagent
- Safe – allows one to perform non-radioactive assays
Cell lysis buffer, reaction buffer and substrate
Renilla luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers.
- Promoter studies for analyzing cis- regulatory elements and trans-acting factors
- Drug screening
- siRNA and miRNA screening
- Multiplexed assays to study off-target effects
- Protein localization reporter assays
- Signal transduction pathway analysis
- RNA splicing studies
The Green Renilla luciferase is a 36kDa protein produced by a derivative of the wild type Renilla luciferase gene from the sea pansy, Renilla reniformis. Compared to the wild type luciferase, Green Renilla is more stable in serum and has an the emission spectrum that is shifted toward the green region. The protein provides extremely bright flash signal that decays rapidly. The Pierce Renilla Flash Assay Kit reagent optimizes the this signal for use in flash-type luciferase reporter assays. The kit also is compatible for flash assays with other Renilla luciferases and derivatives that use coelenterazine as a substrate.
||Renilla luciferase reaction. Coelenterazine is oxidized by Renilla luciferase to produce coelenteramide, carbon dioxide, and light.
||Thermo Scientific Pierce Renilla Luciferase Flash Assay delivers high signal intensity. HEK293 cells were transfected with Thermo Scientific pCMV-Cypridina Luc or the competitor's firefly expression vector. At 24 hours post-transfection, respective flash assay reagents were added to sample wells containing cell lysates.Total light output (RLU) was captured by Thermo Scientific Varioskan Flash Multimode Reader (integration time = 0.1 sec/reading).
- Lorenz, W.W., et al. (1991). Isolation and expression of a cDNA encoding Renilla reniformis luciferase. Proc. Natl. Acad. Sci. USA, 88:4438-4442.
Review of Luciferase Reporters
Luciferase Reporter Assay Selection Guide
Renilla Vectors, Glow and Dual Assay Kits
All Luciferase Reporter Assay Reagents and Kits
Pierce Luciferase Cell Lysis Buffer (2X)
TurboFect in vitro Transfection Reagent
Modified Dulbecco’s PBS BupH Packs
Modified Dulbecco’s PBS, 20X Solution