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

Pierce™ Gaussia-Firefly Luciferase Dual Assay Kit

The Thermo Scientific Pierce Gaussia-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Gaussia and RedRead more
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Catalog NumberQuantity
161821000 Reaction Kit
16181100 Reaction Kit
Catalog number 16182
Price (USD)
1,076.00
Each
Add to cart
Quantity:
1000 Reaction Kit
Price (USD)
1,076.00
Each
Add to cart
The Thermo Scientific Pierce Gaussia-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Gaussia and Red Firefly luciferase activity in mammalian whole cell lysates.

Features of the Gaussia-Firefly Luciferase Dual Assay Kit:

Simultaneous—concurrent, filter-based, wavelength-separated detection of two luciferase activities
Sensitive—measure bright blue Gaussia 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 Gaussia-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Gaussia and Red Firefly luciferase activity. Gaussia 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.

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

Requires:
Gaussia and Red Firefly luciferase reporters; Filter sets in the ranges of 425 to 525nm for Gaussia 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:
Gaussia luciferase produces brighter bioluminescent signal than firefly and native Renilla luciferases. The bioluminescent signal (λmax= 470nm) produced by Gaussia luciferase results from the oxidation of coelenterazine. 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 Gaussia luciferases are used in the dual assay because the light output is spectrally resolvable.

More Product Data
Highly sensitive multiplex luciferase reporter assays
Simultaneous dual-emission detection of luciferase reporter assays
Activation of the antioxidant response pathway by pesticide chemicals
For Research Use Only. Not for use in diagnostic procedures.
Specifications
TypeAssay Kit
TargetLuciferase, Firefly Luciferase, Gaussia Luciferase
Quantity1000 Reaction Kit
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
Substrate PropertiesChemical Substrate
TechniqueEnhanced Chemiluminescence
Substrate TypeLuciferase Substrate
Product LinePierce™
SubstrateCoelenterazine, D-Luciferin
Unit SizeEach
Contents & Storage
• Gaussia-Firefly Dual Assay Buffer, 50 mL, store at 4°C
• Coelenterazine (100X), 0.5 mL, store at -80°C
• D-Luciferin, Lyophilized, 30 mg, store at 4°C
• 2X Cell Lysis Buffer, 60 mL, store at room temperature

Upon receipt store kit at -80°C or store individual components as indicated above.

Frequently asked questions (FAQs)

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.

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

    Here are possible causes and solutions:

    - Low luciferase expression: Lyse cells in smaller volume of 1X Cell Lysis Buffer; Use a different promoter or growth conditions to improve expression; Increase the integration time on the instrument; Scale up volume of sample and reagent per well.

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

    I did not get any signal when I used a Pierce Gaussia/Cypridina/Renilla-Firefly Dual Assay Kit. Why is this?

    Here are possible causes and solutions:

    - Low transfection efficiency: Optimize transfection conditions using a visual transfection control (e.g., a plasmid over-expressing a fluorescent protein); Verify plasmid DNA quality using restriction digestion and agarose gel electrophoresis; Use only transfection grade DNA - please note that most high-quality plasmid DNA should be supercoiled; Use actively dividing, low-passage cells; Use a different cell type.
    - No promoter induction: Incubate cells using promoter-specific inducing conditions; Incubate cells for a longer time after treatment; Change growth conditions to improve expression; Use a different promoter.
    - Substrate auto-oxidized: Protect substrate from light and air; Maintain 100X Coelenterazine at -80 degrees C, 100X Vargulin at -20 degrees C, and 100X D-Luciferin at -20 degrees C.

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

    I got a high background signal with a Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay. What went wrong?

    Here are possible causes and solutions:

    - Non-specific oxidation of substrate: Use less serum in the cell culture media; Note: Albumin can increase the auto-oxidation of Vargulin; Avoid repeated freezing and thawing of the sample.
    - Control sample is contaminated: Use new sample; Change pipette tips after each well.

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