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Amplite® Renilla Luciferase Reporter Gene Assay Kit *Maximized Luminescence*

Renilla Luciferase was measured with Amplite™ Renilla Luciferase Reporter Gene Assay Kit in a white 96-well plate with a NOVOstar plate reader (BMG Labtech). 
Renilla Luciferase was measured with Amplite™ Renilla Luciferase Reporter Gene Assay Kit in a white 96-well plate with a NOVOstar plate reader (BMG Labtech). 
Renilla Luciferase was measured with Amplite™ Renilla Luciferase Reporter Gene Assay Kit in a white 96-well plate with a NOVOstar plate reader (BMG Labtech). 
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Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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ShippingStandard overnight for United States, inquire for international
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Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC41105331

OverviewpdfSDSpdfProtocol


Common reporter genes include beta-galactosidase, beta-glucuronidase and luciferase. The most versatile reporter gene is the firefly luciferase. Recently there is steadily increasing use of other luciferases, such as Renilla luciferase since these reporters are smaller and do not require the presence of ATP. Our Amplite® Renilla Luciferase Reporter Gene Assay Kit is designed to provide a fast and sensitive method to detect the luciferase from sea pansy (Renilla reniformis). It uses a proprietary luminogenic formulation to quantify Renilla luciferase activity in cell-based assays. Our formulation generates a luminescent product that gives strong luminescence upon interaction with Renilla luciferase. The kit provides all the essential components. It has high sensitivity and can be performed in a convenient 96-well and 384-well microtiter-plate format. The "glow-type" signal with a half-life of one hour provides a consistent signal across large number of assay plates. The assay is compatible with standard cell growth media. This kit enables the measurement of primary expression or gene expression with wild type and the synthetic hRluc genes .

Platform


Luminescence microplate reader

Recommended plateSolid white

Components


Example protocol


AT A GLANCE

Protocol summary

  1. Prepare cell plates
  2. Treat cells as needed
  3. Remove medium from cell plates
  4. Add Renilla Luciferase working solution (100 µL/well for 96-well plate or 25 µL/well for 384-well plate)
  5. Incubate at room temperature for 5 - 10 minutes
  6. Monitor luminescence intensity

Important notes
Thaw all the kit components to room temperature before use. For all luminescent experiments, it is recommended to use white plates to get the best results.

PREPARATION OF WORKING SOLUTION

1. Add one volume of 100X Luciferase Substrate (Component A) to 100 volumes of Assay Buffer (Component B) to make Renilla Luciferase working solution. Note: The reconstituted Renilla Luciferase working solution is very sensitive to light, should be kept from light. In addition, it is not stable, should be prepared fresh, kept on ice and used within 2 hours.


For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. Treat cells (or samples) with test compounds by adding 10 µL of 10X test compounds (96-well plate) or 5 µL of 5X test compounds (384-well plate) in desired compound buffer.
     
  2. Incubate the cell plate in a 5% CO2 incubator at 37°C for desired period of time, typically 4 hours to overnight.

  3. Remove the medium completely.

  4. Add 100 µL (96-well plate) or 25 µL (384-well plate) per well of Renilla Luciferase working solution.

  5. Incubate the plate at room temperature for 5 - 10 minutes. Protect from light.

  6. Monitor luminescence intensity with a luminometer.

 

Images


References


View all 56 references: Citation Explorer
RNA detection using peptide-inserted Renilla luciferase
Authors: Andou T, Endoh T, Mie M, Kobatake E.
Journal: Anal Bioanal Chem (2009): 661
The cAMP-dependent protein kinase inhibitor H-89 attenuates the bioluminescence signal produced by Renilla Luciferase
Authors: Herbst KJ, Allen MD, Zhang J.
Journal: PLoS One (2009): e5642
Reassembly of a bioluminescent protein Renilla luciferase directed through DNA hybridization
Authors: Cissell KA, Rahimi Y, Shrestha S, Deo SK.
Journal: Bioconjug Chem (2009): 15
Bioluminescent indicators for Ca2+ based on split Renilla luciferase complementation in living cells
Authors: Kaihara A, Umezawa Y, Furukawa T.
Journal: Anal Sci (2008): 1405
Coelenterazine-binding protein of Renilla muelleri: cDNA cloning, overexpression, and characterization as a substrate of luciferase
Authors: Titushin MS, Markova SV, Frank LA, Malikova NP, Stepanyuk GA, Lee J, Vysotski ES.
Journal: Photochem Photobiol Sci (2008): 189
Structure-function studies on the active site of the coelenterazine-dependent luciferase from Renilla
Authors: Woo J, Howell MH, von Arnim AG.
Journal: Protein Sci (2008): 725
Mutational optimization of the coelenterazine-dependent luciferase from Renilla
Authors: Woo J, von Arnim AG.
Journal: Plant Methods (2008): 23
Red-shifted Renilla reniformis luciferase variants for imaging in living subjects
Authors: Loening AM, Wu AM, Gambhir SS.
Journal: Nat Methods (2007): 641
Crystal structures of the luciferase and green fluorescent protein from Renilla reniformis
Authors: Loening AM, Fenn TD, Gambhir SS.
Journal: J Mol Biol (2007): 1017
Quantification of dynamic protein complexes using Renilla luciferase fragment complementation applied to protein kinase A activities in vivo
Authors: Stefan E, Aquin S, Berger N, L and ry CR, Nyfeler B, Bouvier M, Michnick SW.
Journal: Proc Natl Acad Sci U S A (2007): 16916