Cell Signaling Firefly Luciferase Reporter Systems ONE-Glo™ Luciferase Assay System Product ONE-Glo™ Luciferase Assay System For Laboratory Use. Description: The ONE-Glo™ Luciferase Assay System provides a highly sensitive, robust, homogeneous assay for detection of firefly luciferase reporter gene expression in mammalian cells. Ideally suited for high- and ultrahighthroughput applications, the ONE-Glo™ Assay contains a new luciferase substrate, resulting in a reagent that is more stable, more tolerant to sample components, and has less odor than standard luciferase assay reagents. These features ensure that the ONE-Glo™ Assay provides robust performance and also eliminates many of the handling inconveniences experienced using other reporter assays in a high-throughput setting. Features: • Simplify Your Assay Optimization: The robust performance, reduced odor, improved storage, and larger available sizes make assay optimization even easier and more efficient. • Store at Room Temperature or at 4°C: The extended stability of the ONE-Glo™ Reagent at room temperature and 4°C makes it more convenient for everyday use. • Improve Assay Precision: Because the ONE-Glo™ Reagent is less sensitive to mixing and dispensing conditions, reproducibility is enhanced. The assay is ideal for use in high-density (384- and 1536-well) microplates. • Get a Brighter, Longer-Lasting Signal: Optimized for batch and continuous-process handling, the extended bright light output of the ONEGlo™ Assay allows high sensitivity, especially when extended incubation is required prior to reading results. • Reduce Unwanted Effects from Sample Components: The novel chemistry used in the ONE-Glo™ Assay is less sensitive to culture media, phenol red, and luciferase inhibitors than other luciferase assays. Storage Conditions: Store the ONE-Glo™ Luciferase Assay System components at –20°C. Please refer to the Technical Manual for other storage options, including room-temperature storage. Protocol ONE-Glo™ Luciferase Assay System Technical Manual Part# TM292 1 × 107 Bright-Glo™ Reagent ONE-Glo™ Reagent Steady-Glo® Reagent 1 × 106 ONE-Glo™ Reagent is more tolerant of phenol red than luciferinbased reagents. Luciferase reactions composed of 14.9ng/ml luciferase in phenol red-free RPMI medium (with 0.1% Prionex®) and ONE-Glo™ Reagent or another bright-type reagent were initiated in the presence of varied amounts of phenol red. Relative luminescence is the luminescence of reactions containing phenol red/ luminescence from reactions without phenol red x 100; n = 3. 100 120 20 40 60 80 0 05 10 Phenol Red (mg/L) 15 20 Size Cat.# Price (Fr) 10 ml E6110 190.00 100 ml E6120 1330.00 1 L E6130 Pls. Enq. 100 20 40 60 80 0 ONE-Glo™ Reagent Competitor (Brite reagent) ONE-Glo™ Reagent protects the luciferase reaction in the presence of resveratrol, a known luciferase inhibitor (Bakhtiarova, A. et al. (2006) Biochem. Biophys. Res. Comm. 351, 481–4). Luciferase reactions generated by ONE-Glo™ Reagent or another bright-type reagent were initiated in the presence or absence of 10μM resveratrol. Luminescence was initiated and measured by the method noted in the figure above. The relative luminescence is the luminescence from reactions containing resveratrol/ luminescence from reactions without resveratrol x 100; n = 3. ONE-Glo™ Reagent Bright-type 15 1 × 105 020406080 100 120 Time (minutes) ONE-Glo™ Reagent generates bright and stable luminescence that can easily be measured for multiple hours. Samples in 96-well plates consisted of 50μl of purified firefly luciferase (14.9ng/ml with 0.1% Prionex®) combined with 50μl of the respective reagent. Luminescence was measured (1.0 second integration/well) at 3 minutes and periodically for almost 2 hours. All coefficients of variation were < 3%; n = 3. For complete and up-to-date product information visit: www.promega.com/catalog 263 Reporter Assays and Transfection Luminescence (RLU) 6837MA Relative Luminescence (%) Relative Luminescence (%) 6841MA 6840MB Pagina 266
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