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Assay Method Information

Assay Name:  In Vitro TK Inhibition Assay
Description:  Colorimetric cell proliferation and viability assay (reagent CellTiter-Blue purchased from Promega cat NG8081) was performed on BaF3 Kit WT cell line.A total of 2.104 cells/50 μl were seeded per well of a 96-wells plate. Treatment was initiated by addition of a 2 drug solution of serial dilutions ranging from 0 to 10 μM. After incubating for 48 hours at 37 C., 10 μl of a dilution of CellTiter-Blue reagent was added to each well and the plates were returned to the incubator for an additional 4 hours. The fluorescence intensity from the CellTiter-Blue reagent is proportional to the number of viable cells and data were recorded (544Ex/590Em) using a POLARstar OMEGA microplate reader (BMG LabteckSarl). A background control without cells was used as a blank. The positive control of the assay corresponds to the cell proliferation obtained in the absence of drug treatment (100% proliferation). Each sample was done in triplicate. The results were expressed as a percentage of the proliferation obtained in absence of treatment. All drugs were prepared as 20 mM stock solutions in DMSO and conserved at −80 C. Drug dilutions were made fresh in medium before each experiment. A DMSO control was included in each experiment.
Affinity data for this assay
 

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Last update November 1, 2007
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