BioLux® Gaussia Luciferase Assay Kit
The BioLux® Gaussia Luciferase Assay Kit contains the reagents necessary for assaying Gaussia luciferase (GLuc) activity in the culture media or cell lysate
- The standard assay gives enhanced light output
- The stabilized assay can be used in high-throughput format
- The enhanced signal stability can be adjusted with the desired amount of stabilizer
- The light emission in a stabilized GLuc assay decays slowly with a half-life of approximately 25 minutes
The BioLux® Gaussia Luciferase Assay Kit contains the reagents necessary for assaying Gaussia Luciferase (GLuc) activity, most commonly from cell culture supernatants. Gaussia Luciferase is a reporter luciferase from the marine copepod Gaussia princeps (1,2). Gaussia Luciferase can be expressed in mammalian cells using reporter plasmids available from NEB (Refer to the Companion Products). This luciferase, which does not require ATP, catalyzes the oxidation of the substrate coelenterazine in a reaction that produces light (Figure 1), and has considerable advantages over other luminescent reporter genes.
This kit now includes an additional stabilizer component, which allows the use of the assay in high throughput format or without the requirement of an injector-equipped luminometer. This three-component assay system provides the user with 2 options: (a) use the assay without stabilizer for enhanced light output or (b) use with the desired amount of stabilizer for enhanced signal stability. The stabilizer component allows the use of the assay in high throughput format or without the requirement of an injector-luminometer. For standard assays giving the highest activity, the kit can be used with the GLuc substrate mixed in the assay buffer. With the stabilized assay protocols, the light emission decays slowly with a half-life of approximately 25 minutes. The addition of stabilizer decreases the absolute value of light output but confers signal stability over time (Figure 2).
The luminescence measured from the supernatant of cultured cells transfected with a plasmid expressing GLuc is proportional to the amount of enzyme produced, which in turn, reflects the level of transcription. Alternatively, a cell lysate sample can be used for the assay. Although most of the GLuc is secreted, the high sensitivity of GLuc allows measurements from the cellular fraction.
The following reagents are supplied with this product:
Store at (°C) Concentration BioLux® GLuc Assay Buffer -20 1 X BioLux® GLuc Substrate -20 100 X BioLux® GLuc Stabilizer -20 100 X
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