Figure 1. CellTox™ Green Dye binds DNA of cells with impaired membrane integrity. Viable Cell Low Fluorescence Nonviable Cell High Fluorescence Multiplexing assays gathers more than one set of data from the same sample. The key benefit of multiplexing is gaining a better understanding of the event you are measuring in the context of another parameter, minimizing faulty interpretation of data or ambiguity from data sets. Multiplexing insures that the variables for a sample are the same for all the assays and new variables are not introduced through replicate plating. Multiplexing also provides a way to normalize data to a particular parameter; for instance, coupling the ratio of an activity to the number of cells in the well. To multiplex two or more assays, the assays must meet certain criteria: the signals for the various assays must be spectrally or temporally distinct, the assay chemistries must be compatible, and the assays must fit into the same well or be easily separated (e.g., moving medium to a separate plate and performing further experiments on the cells). CellTiter-Fluor™ Cell Viability Assay CellTiter-Fluor™ Cell Viability Assay + CellTox™ Green Cytotoxicity Assay CellTox™ Green Cytotoxicity Assay A. 100 200 300 400 500 0 IC50 = 1.67–2.05 × 105 2.0 × 105 1.5 × 105 IC50 = 1.64–2.10 × 105 1.0 × 105 0.5 × 105 –6 Log10 –5 [terfenadine], M –4 0 B. 100 200 300 400 500 0 IC50 = 5.50–7.82 × 106 IC50 = 5.13–7.82 × 106 2.0 × 104 4.0 × 104 6.0 × 104 8.0 × 104 –6 Log10 –5 [ionomycin], M Figure 2. Multiplexing the CellTiter-Fluor™ Cell Viability Assay with the CellTox™ Green Cytotoxicity Assay. HEK 293 cells were treated for 6 hours with a serial dilution of terfenadine (Panel A) or ionomycin (Panel B) and tested for viability and cytotoxicity. CellTox™ Green Dye was incorporated into the medium (no-step method) for half of the samples (+ CellTox™ Green Cytotoxicity Assay). Right axis represents signal from CellTox™ Green Dye and left axis represent signal from CellTiter-Fluor™ Viability Assay, both in relative fluorescent units (RFU). Each point and standard deviation represents six replicates. IC50 Read the complete PubHub Article » Learn More enews | April 2015 –4 0 values are listed for samples with and without CellTox™ Green Dye. Viability Fluorescence (RFU) Cytotoxicity Fluorescence (RFU) Viability Fluorescence (RFU) 10887MA Cytotoxicity Fluorescence (RFU) 11698MA Control Control Pagina 8
Pagina 10Interactieve digi-reisgids, deze maandblad of editie is levensecht online geplaatst met Online Touch en bied het digitaal bladerbaar uitgeven van edrukwerk.
Promega Benelux - Monthly online Magazine - April 2015 edition Lees publicatie 100Home