Promega Corporation

Multiplexing Cell-Based Assays Using Standard Luminescent and Fluorescent Plate Readers...

Multiplexing Cell-Based Assays Using Standard Luminescent and Fluorescent Plate Readers Scientific Poster

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Abstract

Richard A. Moravec, Andrew L. Niles, Terry L. Riss, James J. Cali, Peter F. Stecha, Brian D. Almond and Erika M. Hawkins

We have developed methods to combine fluorescent and luminescent cell-based assays to measure more than one parameter from a single sample using ordinary microplate readers. Measuring two or more parameters from the same sample of cells is more efficient, improves consistency compared to using parallel samples, and provides additional information about cellular response. By understanding the assay chemistries and detection parameters, many different assays can be combined in a simultaneous, sequential, or split sample format. We have been successful in multiplexing simultaneous homogeneous fluorescent and luminescent assays for cell viability, caspase activity and total cell number in the same well. Additional homogeneous multiplex examples include real time Renilla luciferase reporter gene assays followed by measurement of cell viability or caspase activity. We can overcome assay chemistry incompatibility for situations such as such as luminescent cytochrome P450 and luminescent cell viability assays by splitting the sample and transferring cell culture supernatant to a separate well. While each assay has individually been proven to be successful in a high throughput format, we have continued to improve protocols that enable multiplex detection of more than one indicator from a single sample.

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  • Part# PS014
  • Printed in USA.

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