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

Pierce™ Renilla Luciferase Flash Assay Kit

The Thermo Scientific Pierce Renilla Luciferase Flash Assay Kit provides researchers with a highly sensitive intracellular assay for transcriptional activityRead more
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Catalog NumberQuantity
161651000 Reaction Kit
16164100 Reaction Kit
Catalog number 16165
Price (CNY)
7,157.00
Each
Add to cart
Quantity:
1000 Reaction Kit
Price (CNY)
7,157.00
Each
Add to cart

The Thermo Scientific Pierce Renilla Luciferase Flash Assay Kit provides researchers with a highly sensitive intracellular assay for transcriptional activity of regulatory elements in mammalian whole cell lysate.

Features of the Renilla Luciferase Flash Assay Kit:

  • Sensitive—greater sensitivity allows utilization of smaller numbers of cells
  • Cost effective—highest sensitivity assays result in decreased reagent consumption
  • Time-saving—assays 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 Renilla luciferase in mammalian cell lysates. When used with Thermo Scientific Green Renilla Luc Plasmids, the kit provides an extremely sensitive bioluminescent reporter assay system for intracellular detection of promoter or pathway activity. The signal produced by Green Renilla Luc is considerably greater than signal from either native firefly or Renilla luciferases assayed under similar conditions. Light output captured using a luminometer can be correlated with the amount of Renilla luciferase protein produced and used to determine the activity of the promoter driving Renilla expression.

Includes:

Cell lysis buffer, reaction buffer and substrate

Requires:

Renilla luciferase and luminometer or other instrument capable of monitoring luminescence, such as the Varioskan ALF or the Varioskan LUX 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 Flash Assay Kit reagent optimizes the this signal for use in flash-type luciferase reporter assays. The kit also is compatible for flash assays with other Renilla luciferases and derivatives that use coelenterazine as a substrate.

More Product Data

Related Products

Pierce™ Renilla-Firefly Luciferase Dual Assay Kit

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
Quantity1000 Reaction Kit
SubstrateCoelenterazine
Substrate PropertiesChemical Substrate
Substrate TypeLuciferase Substrate
TargetLuciferase, Green Renilla Luciferase
TechniqueEnhanced Chemiluminescence
TypeAssay Kit
Unit SizeEach
Contents & Storage
• Renilla Flash Assay Buffer, 50 mL, store at 4°C
• Coelenterazine (100X), 0.5 mL, store at -80°C
• 2X Cell Lysis Buffer, 60 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.

    View all Pierce™ Renilla Luciferase Flash Assay Kit, 1000 Reaction Kit FAQs