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

Ordering Information

$name
  • 100 assays
    $106.00
  • 1,000 assays
    $628.00
  • Product Information
    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).

    Figure 1: The photochemical reaction catalyzed by Cypridina Luciferase. Figure 1: The photochemical reaction catalyzed by Cypridina Luciferase.

    Figure 2: Activity of secreted CLuc at different time points. Figure 2: Activity of secreted CLuc at different time points.

    Supernatants of transfected cells were collected and medium was replaced each day for 4 days. These supernatants were stored at -20°C until the last samples were obtained. Relative Light Units, RLU.
    Figure 3: Sensitivity of Cypridina Luciferase assay. Figure 3: Sensitivity of Cypridina Luciferase assay.

    The CLuc activity was assayed from 10-fold serial dilutions of the supernatant from a stable CLuc-expressing cell line.
    Figure 4: Stability of Cypridina Luciferase. Figure 4: Stability of Cypridina Luciferase.

    (A) Supernatant obtained from stable CLuc-expressing cells was incubated at 55°C or 95°C for 30 minutes and allowed to cool to room temperature (25°C). Five-fold serial dilutions of supernatant were assayed for CLuc activity. The control is the supernatant of the parental cells from which the stable cell line was created. (B) Supernatants from stable CLuc-expressing cells, grown in medium with or without β-mercaptoethanol, were incubated at 37°C and assayed each day for 7 days.
    Figure 5: Kinetics of light emission. Figure 5: Kinetics of light emission.

    The Cypridina Luciferase kinetic assay was performed with the supernatant from HeLa cells transiently
    transfected with a CLuc-expressing vector. The assay solution was prepared and incubated at room
    temperature for 30 minutes before use.
    Figure 6: Cypridina Luciferase Assays Figure 6: Cypridina Luciferase Assays

    Figure 7: Activity of Cypridina Luciferase in the supernatant and lysate from a stable CLuc-expressing cell line. Figure 7: Activity of Cypridina Luciferase in the supernatant and lysate from a stable CLuc-expressing cell line

    The CLuc activity was measured from 20 μl of supernatant (from 500 μl total volume) and from 20 μl of cell lysate (100 μl total lysate volume) of a stable CLuc-expressing cell line.

    Kit Components

    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
    Product Categories:
    Cypridina Luciferase Products
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    Cypridina Luciferase
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