BioLux® Cypridina Luciferase Assay Kit
The BioLux® Cypridina Luciferase Assay Kit contains the reagents necessary for assaying Cypridina Luciferase (CLuc) activity.
- Cypridina luciferase (CLuc) is secreted from cells by virtue of its natural signal peptide and its luminescence can easily be measured from the growth media of transfected cells
- Can be used to assay CLuc activity in the culture media or cell lysate
- CLuc does not cross-react with other luciferases expressed in the same cells
The BioLux Cypridina Luciferase Assay Kit contains the reagents necessary for assaying Cypridina Luciferase activity. Cypridina is a new secreted luciferase reporter from the ostracod Cypridina noctiluca. This luciferase does not require ATP and catalyzes the oxidation of its luciferin substrate in a photochemical reaction (1,2) (Figure 1). The substrate for Cypridina is different than coelenterazine, which is the common substrate of other marine luciferases including Renilla and Gaussia.
Properties of Cypridina Luciferase:
Cypridina Luciferase is secreted from cells by virtue of its natural signal peptide and its luminescence can be measured from the supernatant of transfected cells. Therefore, cell lysis is not necessary (Figure 2).
Secreted CLuc is a very stable protein. Because of this property, the activity measured from the supernatant reflects the amount of protein accumulated up to the time of sampling. Multiple samples can therefore be obtained from the same transfected cells (Figure 2).
The CLuc assay is very sensitive, allowing detection of very small amounts of Cypridina Luciferase activity (Figure 3).
Secreted CLuc is thermally stable at 55°C (Figure 4A), which is the typical inactivation temperature of most viruses. Secreted CLuc remains active in the presence of β-mercaptoethanol, which is commonly present in the complete culture medium of mouse ES cells (Figure 4B).
Although the light reaction catalyzed by Cypridina has an initial emission half-life of approximately 5 minutes, light production continues to decay slowly, and is readily detectable 25 minutes after substrate addition (Figure 5).
The 0.5 ml BioLux Cypridina Luciferase Substrate Solvent must be completely thawed and diluted with absolute ethanol (not included) before using it to make the 100X substrate solution (please refer to the section of Reconstitution of BioLux Cypridina Luciferase Substrate).
The following reagents are supplied with this product:
Store at (°C) Concentration BioLux Cluc Assay Buffer -20 1 X BioLux Cluc Substrate Solvent -20 lyophilized BioLux Cluc Substrate -20
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