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Pierce Cypridina-Firefly Luciferase Dual Assay Kit


Detect two spectrally separated luciferase reporters in a single sample.

Pierce Cypridina-Firefly Dual Luciferase Assay Kit

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. As shown in the animation videos below, 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.

Applications:

  • 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)

Application Videos:

Perform a dual-spectral luciferase assay Perform a time-course assay with secreted luciferases

Product Details:

Cypridina luciferase produces brighter bioluminescent signal than firefly and native Renilla luciferases. The bioluminescent signal (λmax=463nm) produced by Cypridina luciferase results from the oxidation of Vargulin. This reaction does not require adenosine triphosphate (ATP) or other cofactors. Red firefly luciferase is a mutant form of the Japanese firefly luciferase from Luciola cruciata. This luciferase produces a red-shifted emission spectrum (λmax= 613nm) that results from the oxidation of D-luciferin in the presence of ATP. Red firefly and Cypridina luciferases are used in the dual assay because the signals are spectrally resolvable.

Pierce Cypridina-Firefly Dual Luciferase Assay spectra
Thermo Scientific Pierce Cypridina-Firefly Luciferase Dual Assay emission spectra profile. Shaded ranges represent the filter sets used in product development experiments: 480±20nm band pass (BP) and 640nm long pass (LP).

 

Pierce Cypridina-Firefly Dual Luciferase Reporter Assay protocol
Protocol summary for the Thermo Scientific Pierce Cypridina-Firefly Luciferase Dual Assay Kit. This is a one-step procedure for spectrally resolving two luciferases. By including both substrates in a single assay reagent, two luciferase activities can be measured simultaneously in a sample by filter-based detection.

 

More Product Data

General References:

  1. Shimomura, O. (2006). Bioluminescence: Chemical Principles and Methods. World Scientific Publishing Co. Pte. Ltd, Hakensack NJ.

Related Resources:

Review of Luciferase Reporters
Luciferase Reporter Assay Selection Guide
Dual-spectral Luciferase Luminescence Calculator

Related Products:

Cypridina Vectors, Flash and Glow Assay Kits
Firefly Vectors, Flash and Glow Assay Kits
All Luciferase Reporter Assay Reagents and Kits
Pierce Luciferase Cell Lysis Buffer (2X)
TurboFect in vitro Transfection Reagent
Modified Dulbecco’s PBS BupH Packs
Modified Dulbecco’s PBS, 20X Solution


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
16183 Pierce Cypridina-Firefly Luciferase Dual Assay Kit
Formulation: Four-component kit
Sufficient For: 100 dual-luciferase assays in microplate wells

Kit Contents:
Cypridina-Firefly Dual Assay Buffer, 5mL
100X Vargulin, 50┬ÁL
D-Luciferin, 3mg
Luciferase Cell Lysis Buffer (2X), 6mL
100-rxn kit Product Instructions for product #16183 Pierce Cypridina-Firefly Luciferase Dual Assay Kit MSDS for product #16183 Pierce Cypridina-Firefly Luciferase Dual Assay Kit Certificate of Analysis for product #16183 Pierce Cypridina-Firefly Luciferase Dual Assay Kit $99.00

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16184 Pierce Cypridina-Firefly Luciferase Dual Assay Kit
Formulation: Four-component kit
Sufficient For: 1000 dual-luciferase assays in microplate wells

Kit Contents:
Cypridina-Firefly Dual Assay Buffer, 50mL
100X Vargulin, 0.5mL
D-Luciferin, 30mg
Luciferase Cell Lysis Buffer (2X), 60mL
1000-rxn kit Product Instructions for product #16184 Pierce Cypridina-Firefly Luciferase Dual Assay Kit MSDS for product #16184 Pierce Cypridina-Firefly Luciferase Dual Assay Kit Certificate of Analysis for product #16184 Pierce Cypridina-Firefly Luciferase Dual Assay Kit $678.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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