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

Pierce™ Renilla-Firefly Luciferase Dual Assay Kit

The Thermo Scientific Pierce Renilla-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Renilla and RedRead more
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Catalog NumberQuantity
16185100 Reaction Kit
161861000 Reaction Kit
Catalog number 16185
Price (USD)
168.00
Each
In stock
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Quantity:
100 Reaction Kit
Recurring order eligible. Learn more »
Price (USD)
168.00
Each
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The Thermo Scientific Pierce Renilla-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Renilla and Red Firefly luciferase activity in mammalian whole cell lysates.

Features of the Renilla-Firefly Luciferase Dual Assay Kit:

Simultaneous—concurrent, filter-based, wavelength-separated detection of two luciferase activities
Sensitive—measure Green Renilla 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)

The Pierce Renilla-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Green Renilla and Red Firefly luciferase activity. Green Renilla luciferase acts as an experimental reporter with constitutively active red firefly as a normalization control. As shown in the animation video 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.

Includes:
Cell lysis buffer, reaction buffer, and substrates coelenterazine and D-luciferin

Requires:
Green Renilla and Red Firefly luciferase reporters; Filter sets at 525nm±20nm for Green Renilla 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)

Green Renilla produces brighter bioluminescent signal than firefly and native Renilla luciferases. The bioluminescent signal (λmax= 535nm) produced by Green Renilla luciferase protein results from the oxidation of coelenterazine. The second assay uses red firefly luciferase, which is a mutant form of the Japanese firefly luciferase 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.

More Product Data
Simultaneous dual-emission detection of luciferase reporter assays

Related Products
Pierce™ Renilla-Firefly Luciferase Dual Assay Kit
WARNING: Reproductive Harm - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
AssayReporter Enzyme, Luciferase Reporter Assay
Compatible CellsMammalian Cells
Detection MethodBioluminescence
For Use With (Equipment)Luminometer (Microplate)
Format384-well plate, 96-well plate
Label TypeEnzyme Labeled
Product LinePierce™
Quantity100 Reaction Kit
SubstrateCoelenterazine, D-Luciferin
Substrate PropertiesChemical Substrate
Substrate TypeLuciferase Substrate
Sufficient For100 Dual Luciferase Assays
TargetLuciferase, Firefly Luciferase, Green Renilla Luciferase
TechniqueEnhanced Chemiluminescence
TypeAssay Kit
Unit SizeEach
Contents & Storage
Sufficient For: 100 dual-luciferase assays in microplate wells
• Renilla-Firefly Dual Assay Buffer, 5 mL (store at 4°C)
• 100X Coelenterazine, 50 μL (store at -80°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 -80°C, or store individual components as indicated above.
Have questions about this product? Ask our AI assisted search.
I did not get any signal when I used a Pierce Gaussia/Cypridina/Renilla-Firefly Dual Assay Kit. Why is this?
What instrument filters are required to read the luciferase assays?
I am getting a high saturating signal with a Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay. What could have happened?
Which luciferase assays can be used to specifically test media as a sample type?
Why am I getting a low signal in lysate using a Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay? Can you provide some suggestions?
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Frequently asked questions (FAQs)

What are the advantages of using TurboFect Transfection Reagent (Cat. No. R0533) over other transfection reagents with the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186)?

The Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) luciferase vectors were optimized with TurboFect. However, it should be feasible to replace TurboFect with another transfection reagent if your cells tolerate the transfection reagent better and give you acceptable transfection rates.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I store my Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) cell lysates at -20 degrees C to test later?

The stability of the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) may be inconsistent from sample to sample at -20 degrees C. We recommend snap-freezing lysates at -80 degrees C with protease inhibitors to help prevent protein degradation.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I use the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186) with a luminometer that is not equipped with reagent injectors?

No. All Luciferase Dual Assay Kits, such as the Pierce Renilla-Firefly Luciferase Dual Assay Kit (Cat. Nos. 16185, 16186), are flash-based assays. Flash luminescence reactions give a very bright signal over a short reaction time. Luciferase assays that are flash-based require the use of a luminometer or other luminescence-monitoring instrument equipped with automated reagent injectors.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I got a high background signal when I used a Pierce Gaussia/Cypridina/Renilla -Firefly Dual Assay Kit. Why is this?

Here are possible causes and solutions:

- Non-specific oxidation of substrate: Use a new sample; Avoid repeated freezing and thawing of the sample.
- Control sample is contaminated: Change pipette tips after each well; Reduce shaker speed during the cell lysis step to avoid contaminating the wells.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I got a high signal when I used a Pierce Gaussia/Cypridina/Renilla -Firefly Dual Assay Kit. Why is this?

This could be due to high luciferase expression. Here are some suggestions:

  • Reduce incubation time before collecting samples. 
  • Decrease the integration time on the instrument.
  • Dilute the sample: 
    Note: A low sample volume can increase assay variability. Dilute the sample and use the recommended volume of 10-20 µL per assay.



  • Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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