3H-thymidine incorporation assay

US Ulrike Schmidt
GH Gerwin Heller
GT Gerald Timelthaler
PH Petra Heffeter
ZS Zsolt Somodi
NS Norbert Schweifer
MS Maria Sibilia
WB Walter Berger
AC Agnes Csiszar
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Cells were pretreated with different concentrations of crizotinib for 24 h and seeded in triplicates in 96 well plates in the presence of the same inhibitor concentrations. After 24 h of incubation, cells were labeled with 30 μCi/ml methyl 3H-thymidine for 2 h. Radioactive media was removed, cells were washed in phosphate-buffered saline (PBS) and trypsinized. Cells were fixed by Tomtec cell harvester (Tomtec Inc., USA) onto a wax-embedded filtermat, and radioactive intensity was determined by a Wallac 1450 MicroBeta liquid scintillator (PerkinElmer Inc., USA). Results were normalized according to cell number.

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