Pierce™ Gaussia Luciferase Flash Assay Kit
Pierce™ Gaussia Luciferase Flash Assay Kit
Thermo Scientific™

Pierce™ Gaussia Luciferase Flash Assay Kit

The Thermo Scientific Pierce Gaussia Luciferase Flash Assay Kit provides researchers with the most sensitive assay for transcriptional activity ofRead more
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Catalog NumberQuantity
161591000 Reaction Kit
16158100 Reaction Kit
Catalog number 16159
Price (USD)
666.00
Each
Add to cart
Quantity:
1000 Reaction Kit
Price (USD)
666.00
Each
Add to cart
The Thermo Scientific Pierce Gaussia Luciferase Flash Assay Kit provides researchers with the most sensitive assay for transcriptional activity of regulatory elements in mammalian cell culture media and whole cell lysate.

Features of the Gaussia Luciferase Flash Assay Kit:

• Sensitive—1000 times greater sensitivity allows utilization of smaller numbers of cells
• Cost effective—highest sensitivity assays result in decreased reagent consumption
• Secreted luciferase—allows real-time assays and kinetic studies without destroying cells
• Time-saving—assays using secreted luciferases require minimal sample handling
• Automation-friendly—amenable to high throughput screens
• Convenient—contains a universal cell lysis buffer and optimized flash assay reagent
• Safe—allows one to perform non-radioactive assays

This Flash Assay Kit contains reagents for measuring the activity of Gaussia luciferase in mammalian cell culture media and lysates. When used with Thermo Scientific Gaussia Luc Vectors, the kit provides an extremely sensitive bioluminescent reporter assay system for secreted or intracellular detection of promoter or pathway activity. Gaussia luciferase is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity. The signal produced by Gaussia is considerably greater than signal from either firefly or Renilla luciferases assayed under similar conditions.

Includes:
Cell lysis buffer, reaction buffer and substrate

Requires:
Gaussia luciferase and luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers.

Applications:
• Promoter studies for analyzing cis- regulatory elements and trans-acting factors
• Drug screening
• siRNA and miRNA screening
• Multiplexed assays to study off-target effects
• Secretory pathway/protein localization reporter assays
• Signal transduction pathway analysis
• RNA splicing studies

Gaussia luciferase is a 20kDa protein from the marine copepod, Gaussia princeps. The bioluminescent enzyme is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity. Light output generated by the luciferase reaction can be correlated to the amount of Gaussia luciferase protein produced and used to determine the activity of the promoter driving Gaussia expression. The signal produced by Gaussia luciferase shows strong flash kinetics and is considerably greater than flash signals from either firefly or Renilla luciferases assayed under similar conditions. The assay kit is optimized for use with our Gaussia Luc Vectors but is fully compatible with other Gaussia luciferases and derivatives that use coelenterazine as a substrate. (For glow assays, use our Gaussia-Dura Luc Vectors.)

More Product Data
• Highly sensitive multiplex luciferase reporter assays
• Luciferase assays in hard-to-transfect Jurkat cells
• Monitoring neuronal differentiation using multiplexed luciferase reporters
• Activation of the antioxidant response pathway by pesticide chemicals
• Versatile luciferases: microplate luminometers and flash luciferase assays
WARNING: Reproductive Harm - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
TypeAssay Kit
TargetLuciferase, Gaussia Luciferase
Sufficient For1000 Luciferase Assays
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
Unit SizeEach
Contents & Storage
Sufficient For: 1000 luciferase assays in microplate wells
• Gaussia Flash Assay Buffer, 50 mL (store at 4°C)
• 100X Coelenterazine, 0.5 mL (store at -80°C)
• Luciferase Cell Lysis Buffer (2X), 60 mL (store at room temperature)

Kit may be stored at -80°C, or individually at indicated temperatures.

Frequently asked questions (FAQs)

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.

I am getting a high saturating signal with a Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay. What could have happened?

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: for secreted Gaussia/Cypridina Luciferase, dilute the sample using media from the cell culture; for cell lysate, dilute the sample using lysis buffer 


    • 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.

    Why am I getting a low signal in lysate using a Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay? Can you provide some suggestions?

    Here are possible causes and solutions:

    - Non-optimized lysis buffer used: Assay luciferase activity in the media to confirm good expression of luciferase; Use only the provided lysis buffer.
    - 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 got a low signal in media when I used a Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay. What happened?

    Here are possible causes and solutions:

    - Insufficient luciferase accumulation in media: Incubate cells for a longer time.
    - Low luciferase expression: Use less media per well during the experiment;.Use a different promoter or growth conditions to improve expression;.Increase the integration time on the instrument;.Scale-up the volume of sample and reagent per well.
    - Treatment interfered with cellular secretory pathway: Transfect cells with a plasmid for constitutive expression of Luciferase (i.e., with pTK or pCMV promoter); Determine if luciferase actively expresses in media without treatment. Add treatment; Determine if there is a corresponding drop in luciferase activity from the constitutively expressed plasmid.

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

    I am getting no signal with my Gaussia/Cypridina/Renilla Luciferase Glow/Luciferase Flash assay. 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 - use only transfection grade DNA; Use actively dividing, low passage cells; Use a different cell type.
    - No or low promoter activity: Use conditions known for promoter activation; Incubate cells for a longer time; 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; maintain 100X Vargulin at -80 degrees C; Prepare new Coelenterazine Working Solution if used longer than 8 hours; Prepare new Vargulin Working Solution if used longer than 2 hours

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

    View all Pierce™ Gaussia Luciferase Flash Assay Kit FAQs