SuperLight™ Luciferase Reporter Gene Assay Kit

Application

• Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens.

Key Features

• High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude.
• Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems.
• Fast and convenient. Homogeneous “mix-and-measure” assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection.
• Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.

Method

• Luminescence

Samples

• Cells etc

Species

• All

Procedure

• 2 min

Size

• 500 tests

Detection Limit

• 2 fg luciferase

Shelf Life

• 12 months

More Details

• The SuperLight™ Luciferase Reporter Gene Assay is based on the quantitation of luciferase expression in mammalian, yeast or E. coil cells, using luciferin and ATP as substrates. The reaction results in light production which can be conveniently measured on a luminometer. This bioluminescent reporter gene assay is extremely sensitive and is especially suitable for quantifying luciferase expression in recombinant cells. This ultra-sensitive, homogeneous cell-based assay only requires adding a single reagent to the cells and measuring the light intensity after a short incubation step (2 minutes). Assays can be performed in tubes, cuvettes or multi-well plates. All kit components are compatible with culture media and with all liquid handling systems. With an extended luminescence emission kinetics (half-life 40 min), the SuperLight™ luciferase assays are especially suitable for high-throughput screening in 96-well, 384-well and 1536-well plates. In addition, the reagent provided in the kits has been formulated for maximum sensitivity, reproducibility and long shelf-life. Applications for this kit include gene regulation studies and high-throughput screening of gene modulators

Can we determine ATP concentration in E. coli cells?

Yes. Ishida A (2002, Anal. Biochem 305, 236-241) described using equal volumes of 10% TCA and E. coli cell culture. Let stand 3 min at room temp. Mix 1 vol of the cell extract with 19 vol of 25 mM Hepes, 17 mM NaOH, and adjust pH to 7.8.before assaying.

How to adapt your protocol for adherent cells when working in a 24-well format?

Lyse the cells in the 24 well plate, then add 50 µL cell lysate to 50 µL SuperLight Reagent (10 µL + 10 µL would work too). Alternatively, cells could be harvested with Trypsin/EDTA, resuspend cells in the same volume (200-500 µL of medium). Do the 50 µL + 50 µL assay.

Cohen ME, et al (2011). Antibody against extracellular vaccinia virus (EV) protects mice through complement and Fc receptors. PLoS One. 6(6):e20597. Assay: Reporter gene in monkey cell line.

Fuller CA, et al (2011). Shiga toxin subtypes display dramatic differences in potency. Infect Immun. 79(3):1329-37. Assay: Reporter gene in monkey cell line.

Mayer-Wagner S et al (2011). Effects of low frequency electromagnetic fields on the chondrogenic differentiation of human mesenchymal stem cells. Bioelectromagnetics. 32(4):283-90. Assay: Luciferase reporter gene in monkey vero cells.

Tannous BA et al (2011). Gaussia Luciferase Variant For High-Throughput Screening. WO2011002924. Assay: Reporter gene in mice.

Kulkarni AA, et al (2010). Glycan encapsulated gold nanoparticles selectively inhibit shiga toxins 1 and 2. Bioconjug Chem. 21(8):1486-93. Assay: Reporter gene in monkey cell line.

Labuzek K et al (2010). Ambivalent effects of compound C (dorsomorphin) on inflammatory response in LPS-stimulated rat primary microglial cultures. Naunyn Schmiedebergs Arch Pharmacol. 381(1):41-57. Assay: Luciferase reporter gene in human cell line.

Labuzek K et al (2010). Metformin has adenosine-monophosphate activated protein kinase (AMPK)-independent effects on LPS-stimulated rat primary microglial cultures. Pharmacol Rep. 62(5):827-48. Assay: Luciferase reporter gene in human cell line.

McGannon CM, et al (2010). Different classes of antibiotics differentially influence shiga toxin production. Antimicrob Agents Chemother. 54(9):3790-8. Assay: Reporter gene in human cell line.

Roupelieva M, et al (2010). Kaposi’s sarcoma-associated herpesvirus Lana-1 is a major activator of the serum response element and mitogen-activated protein kinase pathways via interactions with the Mediator complex. J Gen Virol. 91(5):1138-49. Assay: Reporter gene in human cell line.

Zaliauskiene L, et al (2010). Efficient gene transfection using novel cationic polymers poly(hydroxyalkylene imines). Bioconjug Chem. 21(9):1602-11. Assay: Reporter gene in mice.

Tannous, BA. et al (2009). Secreted Luciferase For Ex Vivo Monitoring Of In Vivo Precesses. US2009235370. Assay: Reporter gene in mice.

Gentry, M. et al. (2007). Role of Primary human Alveolar Epithelial Cells in Host Defense against Francisella tularensis Infection. Infection and Immunity 75(8): 3969-3978. Assay: Reporter gene in human cell line.

Michael, K. et al. (2007). Microplate Orbital Mixing Improves High-Throughput Cell-Based Reporter Assay Readout. J Biomol Screen 12(1):140-144. Assay: Reporter gene in mouse 3T3 cells.

Saenz JB,et al (2007). Identification and Characterization of Small Molecules That Inhibit Intracellular Toxin Transport. Infection and Immunity 75(9): 4552-4561. Assay: Reporter gene in monkey cell line.

Zhao, L. and Haslam, D.B. (2005). A quantitative and highly sensitive luciferase-based assay for bacterial toxins that inhibit protein synthesis. J Med Microbiol 54:1023-1030. Assay: Reporter gene in human cell line.

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost