The Thermo Scientific Pierce Gaussia Luciferase Flash Assay Kit provides researchers with the most sensitive assay for transcriptional activity of regulatory elements in mammalian cell culture media and whole cell lysate.
This Flash Assay Kit contains reagents for measuring the activity of Gaussia luciferase in mammalian cell culture media and lysates. When used with Thermo Scientific Gaussia Luc Vectors, the kit provides an extremely sensitive bioluminescent reporter assay system for secreted or intracellular detection of promoter or pathway activity. Gaussia luciferase is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity. The signal produced by Gaussia is considerably greater than signal from either firefly or Renilla luciferases assayed under similar conditions.
- Sensitive – 1000 times greater sensitivity allows utilization of smaller numbers of cells
- Cost effective – highest sensitivity assays result in decreased reagent consumption
- Secreted luciferase – allows real-time assays and kinetic studies without destroying cells
- Time-saving – assays using secreted luciferases 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
Gaussia 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
- Secretory pathway / protein localization reporter assays
- Signal transduction pathway analysis
- RNA splicing studies
Gaussia luciferase is a 20kDa protein from the marine copepod, Gaussia princeps. The bioluminescent enzyme is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity. Light output generated by the luciferase reaction can be correlated to the amount of Gaussia luciferase protein produced and used to determine the activity of the promoter driving Gaussia expression. The signal produced by Gaussia luciferase shows strong flash kinetics and is considerably greater than flash signals from either firefly or Renilla luciferases assayed under similar conditions. The assay kit is optimized for use with our Gaussia Luc Vectors but is fully compatible with other Gaussia luciferases and derivatives that use coelenterazine as a substrate. (For glow assays, use our Gaussia-Dura Luc Vectors.)
||Gaussia luciferase reaction. Oxidative decarboxylation of coelenterazine by Gaussia luciferase produces coelenteramide, carbon dioxide, and light.
||Thermo Scientific Pierce Gaussia Luciferase Flash Assay delivers the brightest light intensity. HEK293 cells were transfected with Thermo Scientific pCMV-Gaussia Luc or the competitor's firefly expression vector. At 24 hours post-transfection, respective flash assay reagents were added to sample wells containing either cell media or lysates. Total light output (RLU) was captured by Thermo Scientific Varioskan Flash Multimode Reader (integration time = 0.1 sec/ reading for our assay; = 1sec /reading for the competitor's assay).
|Higher sensitivity for detection in difficult-to-transfect cell lines. Thermo Scientific pCMV-Gaussia plasmid (100ng) was transfected in Jurkat suspension cells by using Thermo Scientific TurboFect Transfection Reagent. Transfection efficiency of <1% was observed by FACS analysis using a CMV-GFP control plasmid. At 24 hours post-transfection, 20µL of sample was used to measure the luciferase activity in each flash assay reagent.
- Szent-Gyorgyi, C., et al. (1999). Cloning and characterization of new bioluminescent proteins. Part of the SPIE Conference on Molecular Imaging: Reporters, Dyes, Markers, and Instrumentation. San Jose, CA. Proc. SPIE 3600:4-11.
- Tannous, B. A., et al. (2005). Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Molecular Therapy 11:435-443.
- Varsano, T., et al. (2013). Inhibition of melanoma growth by small molecules that promote the mitochondrial localization of ATF2. Clin. Cancer Res. 19:2710-22.
Review of Luciferase Reporters
Luciferase Reporter Assay Selection Guide
Gaussia Vectors, Flash and Dual Assay Kits
Gaussia-Dura Vector and Glow Assay Kit
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