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Promegas Multiplexed Luciferase Reporter and Cell Viability Assays performed on the PHERAstar

Tracy Worzella and Brad Larson
Promega Corporation, Madison, WI, USA


Application Note 139 Rev. 7/2006

  • Multiplexing homogeneous cell-based assays for analysis of different parameters from a single sample well
  • Monitoring of reporter gene activity and cell viability
  • Assay miniaturization up to 1536-well format


    • Introduction
    Today's high-throughput screening facilities face increasing demands to generate more information from their existing compound libraries. One method of obtaining this information is to run assays sequentially, looking at one parameter followed by another in different plates. While this option may produce the desired data, the increased time and consumable costs are drawbacks. A more appealing method for data generation is to perform assays in a multiplexed format in which several parameters can be measured within the same well. This multiplexed format not only saves time and consumable cost, but also saves on the useage of valuable test compounds.

    This concept of assay multiplexing is demonstrated here using several cell-based assays multiplexed together. There are inherent properties to cell assays that make them attractive for multiplexed cell-based applications. Cell-based assays are especially vulnerable to variations due to differences in cell growth and metabolism that can arise from plate-to-plate. Cell culture itself is also expensive. By multiplexing assays, fewer cells are needed to acquire the same amount of data. Using the same cells for subsequent assays can also ensure more precise data. In this applicat
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