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Abstract for A Multiplex Viability and Cytotoxicity Assay for
Improving Screening Data
Andrew Niles*, Tracy Worzella*, Richard Moravec*, William Daily#,
Michael Scurria#, Laurent Bernad#, Brian McNamara*, Kay Rashka*,
Deborah Lange*, and Terry Riss*.
*Promega Corporation, Madison, WI 53711 and #Promega Biosciences,
San Luis Obispo, CA 93401
We have developed a homogeneous assay chemistry that
simultaneously measures the relative number of live and dead cells
in culture by detecting changes in cell membrane integrity. This
single
addition assay measures two constitutive proteolytic activities; one
is a marker of viability, and the other a marker of cytotoxicity.
Together, these measures provide an inversely complimentary
viability profile for each assay well. The resulting ratiometric
assay data can also be used to improve additional multiplex endpoint
data quality by response normalization, by increasing content, and
by mitigating false negative or positive determinations. Here we
detail advantages of the assay in simple cytotoxicity screening and
in multiplexed cell-based models of necrosis and apoptosis. The
functionality, sensitivity, and utility of the assay will be
described using high density, multiwell formats with conventional
fluid handling robotics and standard fluorometers or luminometers.
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