â Realizing the potential of IonWorks screening in a drug discovery ...

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Assay format• Single hole HT mode• Pulse to -10mV from -55mV 25times @ 10Hz• First run of day from -100mV– Keep track of relative current– Provides estimate of voltagecontrol• 6 compounds/plate• Full 7-pt IC 50 ’s• One row vehicle control• Four plates/day – 24cmpds/day• 96 cmpds/week
The importance of voltage control onIonWorksNa v Inactivation~ 11mVThe steady-state inactivation curve indicates that an 11mV shift involtage can yield a 50% shift in current amplitude.
Page 1 and 2: Realizing the potential of IonWorks
Page 3 and 4: Groton ion channel drug discoveryCN
Page 5 and 6: Electrophysiology has an integralro
Page 7 and 8: Assessment of cell growth parameter
Page 9 and 10: Growth kinetics dictate experimenta
Page 11: Variables influencing assay success
Page 15 and 16: Sample ion channel recordingPre-com
Page 17 and 18: Assay validation post-relocation•
Page 19 and 20: Resolution approachRe-chlorideE-hea
Page 21 and 22: I-V and inactivation curvesre-visit
Page 23 and 24: Pre- to post-addition current ampli
Page 25 and 26: Summary• Cell line husbandry and
Page 27 and 28: Back-up slides
Page 29 and 30: Kv70Single hole mode70Population pa

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â Realizing the potential of IonWorks screening in a drug discovery ...