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Promegas Multiplexed Cell Viability and Apoptosis Assays performed on the PHERAstar

igure 3). Cell viability and apoptosis were sequentially measured with BMG LABTECHs PHERAstar in both luminescence and fluorescence modes, depending on the multiplex combination. Jurkat cells were treated with various concentrations of anti-FAS monoclonal antibody for 5 hours to induce apoptosis. Cell viability was determined by adding CellTiter-Blue reagent to each well after drug addition and incubating for two hours before recording fluorescence (Ex: 540 nm; Em: 590 nm). The caspase activity was then measured by adding either Caspase-Glo 3/7 or Apo-ONE reagent and incubating for an additional hour prior to recording luminescence or fluorescence (Ex: 485 nm; Em: 520 nm) respectively.

Figures 2 and 3 show the results of two experiments to determine the method of cell death caused by different concentrations of anti-FAS antibody in Jurkat cells. The two experiments measured two different endpoints: reduction of a resazurin dye as an indicator of viable cells and caspase activity as a marker for apoptotic cells. The data show that with increasing concentration of anti-FAS antibody, an increase in caspase-3/7 activity, with a corresponding decrease in cell viability, is observed.



The results suggest that the cell population studied is less viable over the range of treatment due to an increase in apoptosis, as opposed to necrosis.For all cell viability and apoptosis multiplexing combinations, results in 1536 format are comparible to results in 384-well format.


Conclusion
Each of the experiments shown here highlights the ability to perform different assays within the same assay w
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