Cell-based kinase assays in HTS Benjamin Bader

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Lanthascreen – cellular background PTEN negative PI3K E545K • PC3-cells - <strong>in</strong> contrast to MCF7 - have fully stimulated pathway cave different pathway mutations have different impact on basal activation 16 HTRF-symposium 2013 161 16
Lanthascreen - optimization • stimulation time • stimulation temperature • antibody concentration • time cell lysis read • DMSO sensitivity • Volume 10 µl 5 µl • frozen cells 17 HTRF-symposium 2013 171 17
Page 1 and 2: Cell-based kinase assays in HTS Pot
Page 3 and 4: Kinases as drug targets cAbl and Gl
Page 5 and 6: Features of a typical early kinase
Page 7 and 8: General options for cell-based kina
Page 9 and 10: TR-FRET systems: HTRF and Lanthascr
Page 11 and 12: Introducing the PI3K / Akt / mTOR p
Page 13 and 14: Lanthascreen Assay Development BacM
Page 15: Lanthascreen - virus efficiency 30%
Page 19 and 20: Lanthascreen - inhibitor validation
Page 21 and 22: HTRF - cell lines 18000 Ratio 16000
Page 23 and 24: HTRF - pathway inhibitor validation
Page 25 and 26: Focussed medium throughput screen 2
Page 27 and 28: Focussed medium throughput screen L
Page 29 and 30: Acknowledgements Anja Kretzschmar S

Cell-based kinase assays in HTS Benjamin Bader

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