The Thermo Scientific Pierce Cypridina Luciferase Flash Assay Kit provides researchers with a highly 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 Cypridina luciferase in mammalian cell culture media and lysates. When used with Thermo Scientific Cypridina Luc Vectors, the kit provides an extremely sensitive bioluminescent reporter assay system for secreted or intracellular detection of promoter or pathway activity. Cypridina luciferase is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity. The signal produced by Cypridina is considerably greater than signal from either firefly or Renilla luciferases assayed under similar conditions.
- Sensitive – high sensitivity allows utilization of smaller numbers of cells
- Cost effective – sensitive assays result in decreased reagent consumption
- Secreted luciferase – allows real-time assays and kinetic studies without destroying cells
- Ideal for co-transfection – uses a different substrate than Renilla, Gaussia, or firefly luciferases
- Compatible – assay reagents compatible with other Cypridina or Vargula luciferases
- 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
Cypridina 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
The Cypridina luciferase is a 61kDa protein from the marine ostracod, Cypridina noctiluca. Cypridina luciferase is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity. Light output captured using a luminometer can be correlated with the amount of Cypridina luciferase protein produced and used to determine the activity of the promoter driving Cypridina expression. The Cypridina gene in Thermo Scientific Cypridina Luc Vectors is modified for expression in mammalian cells.
The assay kit is optimized for use with our Cypridina Luc Vectors but is fully compatible with any luciferases that use Vargulin as a substrate. Because Cypridina uses a different substrate than Renilla, Gaussia, or firefly luciferases, co-transfection and assays with up to three reporters are possible. The signal produced by Cypridina shows strong flash kinetics and is considerably greater than flash signals from either firefly or Renilla luciferases assayed under similar conditions.
||Cypridina luciferase reaction. Cypridina luciferin, also known as Vargulin, substrate is oxidized by Cypridina luciferase, producing Cypridina oxyluciferin, carbon dioxide and light.
||Thermo Scientific Pierce Cypridina 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 either cell media or lysates. Total light output (RLU) was captured by Thermo Scientific Varioskan Flash Multimode Reader (integration time = 0.1 sec/reading).
- Nakajima, Y., et al. (2004). cDNA Cloning and characterization of a secreted luciferase from the luminous Japanese ostracod, Cypridina noctiluca. Biosci. Biotechnol. Biochem. 68(3):565-570.
- 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
Cypridina 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