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Abstract for A Homogenous Bioluminescent Assay for
High-throughput Antimicrobial Screening
Frank Fan, Braeden Butler, Terry Riss and Keith Wood
Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711
ATP-based detection of microbial cells represents a key
application of luciferase/luciferin bioluminescence assay.
Conventional methods require two steps: application of a lysis
reagent to release microbial ATP, followed by a detection reagent to
elicit bioluminescence. We have developed an assay that combines the
lytic reagent with luciferase/luciferin, thus allowing sensitive
detection of microbial cells in a single-step. The assay system
utilizes a thermostable luciferase to enable extraction of ATP from
bacterial cells and to support a stable “glow-type” luminescent
signal. In addition, we developed a formulation to achieve rapid and
efficient extraction of ATP from a variety of microbial cells. The
combination of these two essential elements enabled the design of a
homogeneous single-reagent system (BacTiter-Glo™ Reagent) for
performing ATP assays on cultured cells.
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