Thermo Scientific™ Pierce™ Gaussia Luciferase Glow Assay Kit

Descripción

The Thermo Scientific™ Pierce Gaussia Luciferase Glow Assay Kit provides an extremely bright and stable bioluminescence signal (half-life greater than 1 hour), especially in the presence of Gaussia-Dura luciferase reporter.

The Thermo Scientific Pierce Gaussia Luciferase Glow Assay Kit provides a sensitive bioluminescence system to determine expression of a gene of interest with a luciferase reporter. The extremely bright and stable bioluminescence signal (half-life greater than 1 hour) is produced in the presence of the Gaussia-Dura luciferase reporter due to oxidation of the luciferase substrate coelenterazine (Figure 1). Gaussia luciferase is a 20 kDa protein from the marine copepod, Gaussia princeps, that has greater protein stability and signal brightness than firefly and native Renilla luciferase. 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.

Features of the Gaussia Luciferase Glow Assay Kit:
• Sensitive—highly sensitive detection of Gaussia luciferase activity
• Stable—increased signal stability
• Simple—does not require a luminometer with injectors
• Compatible—assay reagents compatible with other Gaussia luciferases
• Automation-friendly—amenable to high throughput assays
• Convenient—contains a universal cell lysis buffer and optimized glow assay reagent
• Safe—allows one to perform non-radioactive assays

When used with Thermo Scientific Gaussia-Dura Luc Vectors, the kit provides an extremely sensitive bioluminescent reporter assay system for detection of secreted or intracellular luciferase activity from promoter or pathway activation with extended signal stability. The light output generated by the luciferase reaction (emission maximum of 485 nm) indicates that Gaussia luciferase was transcribed. This light emission can be correlated to the amount of luciferase protein produced, which in turn is proportional to the activity of the promoter for Gaussia luciferase expression and indicates that the regulatory region of the target gene was active. This reaction does not require adenosine triphosphate (ATP) or other cofactors. The kit includes cell lysis buffer, reaction buffer and substrate and can be used to detect the activity of any Gaussia luciferases or derivatives that use coelenterazine as a substrate.

The Gaussia Luciferase Glow Assay Kit requires:
Gaussia luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Varioskan LUX Microplate Readers.

Applications of the Gaussia Luciferase Glow Assay Kit:
• 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 Gaussia-Dura gene used in the Thermo Scientific Gaussia-Dura Luc Vectors is derived from the wild type Gaussia gene and provides equally bright but more stable light output. The Gaussia Glow Assay Kit reagent yields a Gaussia-Dura luciferase reaction with glow-type kinetics that provides a lower intensity but longer-duration signal allowing for higher sample throughput. This glow-type assay with a prolonged luminescence signal does not require reagent injectors and is recommended for luminometers without injectors or for applications requiring batch processing of samples.