Thermo Scientific™ Pierce™ Cypridina-Firefly Luciferase Dual Assay Kit

Simultaneously detect intracellular Cypridina and Red Firefly luciferase activity in mammalian whole cell lysates.

The Thermo Scientific™ Pierce Cypridina-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Cypridina and Red Firefly luciferase activity in mammalian whole cell lysates.

The Pierce Cypridina-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Cypridina and Red Firefly luciferase activity. Cypridina luciferase acts as an experimental reporter with constitutively active red firefly as a normalization control. This reporter-and-control combination enables simultaneous monitoring of experimental reporter and control luciferase activities in a single-read assay without the need for two-step addition of substrate reagents or quenching. The assay working solution contains substrates for both luciferases, and the reactions occur simultaneously with flash-type kinetics. The resulting luminescent signals are spectrally resolvable using filters. In a single sample, researchers can assay transcriptional activity of regulatory elements, signal transduction pathways, and effects of activators or inhibitors.

Highlights:

• Simultaneous – concurrent, filter-based, wavelength-separated detection of two luciferase activities
• Sensitive – measure bright blue Cypridina and red firefly luciferase activities in the same sample
• Fast – no quenching step required, unlike in traditional sequential dual assays
• Multiplex – capable of quantitating two cellular activities in the same sample(s)

Includes:

Cell lysis buffer, reaction buffer, and substrates Vargulin and D-luciferin

Requires:

Cypridina and Red Firefly luciferase reporters; Filter sets in the ranges of 425 to 525nm for Cypridina Luc and 615 to 675nm for Red Firefly Luc. Luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers; Injector required for assessing more than 24 wells at a time.

Recommended for:

Study two regulatory elements at the same time; Monitor two signaling pathways simultaneously; Enable studying more than one target per screen (e.g. off-target effects)

Type

Assay Kit

Target

Luciferase, Cypridina Luciferase

Sufficient For

100 Dual Luciferase Assays

Quantity

100 Reaction Kit

Content And Storage

Sufficient For: 100 dual-luciferase assays in microplate wells
• Cypridina-Firefly Dual Assay Buffer, 5 mL (store at 4°C)
• 100X Vargulin, 50 μL (store at -20°C)
• D-Luciferin, 3 mg (store at 4°C)
• Luciferase Cell Lysis Buffer (2X), 6 mL (store at room temperature)

Upon receipt, store kit at -20°C, or store individual components as indicated above.

Assay

Reporter Enzyme, Luciferase Reporter Assay

Compatible Cells

Mammalian Cells

Detection Method

Bioluminescence

For Use With (Equipment)

Luminometer (Microplate)

Format

384-well plate, 96-well plate

Label Type

Enzyme Labeled

Substrate Properties

Chemical Substrate

Technique

Enhanced Chemiluminescence

Substrate Type

Luciferase Substrate

Product Line

Pierce™

Substrate

Vargulin (Cypridina Luciferin), D-Luciferin

For Research Use Only. Not for use in diagnostic procedures.