SuperLight™ Luciferase Reporter Gene Assay Kit

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.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.

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, pleaseclick here.