Pierce™ Renilla Luciferase Glow Assay Kit
Pierce™ Renilla Luciferase Glow Assay Kit
Thermo Scientific™

Pierce™ Renilla Luciferase Glow Assay Kit

The Thermo Scientific Pierce Renilla Luciferase Glow Assay Kit provides an extremely bright and stable bioluminescence signal (half-life approx. 3Read more
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Catalog NumberQuantity
16166100 Reaction Kit
161671000 Reaction Kit
Catalog number 16166
Price (USD)
132.00
Each
Add to cart
Quantity:
100 Reaction Kit
Price (USD)
132.00
Each
Add to cart
The Thermo Scientific Pierce Renilla Luciferase Glow Assay Kit provides an extremely bright and stable bioluminescence signal (half-life approx. 3 hours), especially in the presence of Green Renilla luciferase reporter.

Features of the Renilla Luciferase Glow Assay Kit:

Sensitive—highly sensitive detection of Green Renilla luciferase activity
Stable—increased signal stability compared to flash assays
Simple—does not require a luminometer with injectors
Compatible—assay reagents compatible with other Renilla luciferases
Automation-friendly—amenable to high throughput assays
Convenient—contains a universal cell lysis buffer and optimized glow assay reagent
Safe—allows one to perform non-radioactive assays

This Pierce Luciferase Glow Assay Kit contains reagents for measuring the activity of Renilla luciferase in cell lysates. In the presence of luciferase, the glow reagent provides stable bioluminescent signal and is highly suited for luminometers without injectors or for batch processing of samples. The light output generated by the luciferase reaction can be correlated with the amount of luciferase protein produced which in turn is proportional to the promoter activity driving the luciferase expression. The glow assay reagents were optimized for best results with Thermo Scientific Green Renilla Luc Plasmids; however, this glow kit may be used to detect the activity of other Renilla luciferase derivatives that use coelenterazine as a substrate.

Includes:
Cell lysis buffer, reaction buffer and substrate

Requires:
Renilla 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
• Protein localization reporter assays
• Signal transduction pathway analysis
• RNA splicing studies

The Green Renilla luciferase is a 36kDa protein produced by a derivative of the wild type Renilla luciferase gene from the sea pansy, Renilla reniformis. Compared to the wild type luciferase, Green Renilla is more stable in serum and has an the emission spectrum that is shifted toward the green region. The protein provides extremely bright flash signal that decays rapidly. The Pierce Renilla Glow Assay Kit reagent effectively stabilizes this bright bioluminescence to yield a signal half-life greater than 3 hours, thus eliminating the need for a luminometer with injectors. The assay kit was optimized for best results with Thermo Scientific Green Renilla Luc Plasmids; however, this glow kit may be used to detect the activity of other Renilla luciferase derivatives that use coelenterazine as a substrate.

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
TypeAssay Kit
TargetLuciferase, Green Renilla Luciferase
Quantity100 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
Upon receipt store kit at -80°C or store individual components as indicated above.
• Renilla Glow Assay Buffer, 5 mL, store at 4°C
• Coelenterazine (100X), 50 μL, store at -80°C
• 2X Cell Lysis Buffer, 6 mL, store at room temperature

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.