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

More documents

Recommendations

Info

Cell preparation affects seal quality and currentamplitudeAverage Seal Resistance350~ 180 MΩ ~ 268 MΩSeal Resistance (MOhms)300250200150100Versene PrepIonWorks HT Experimental RunsVersene / Trypsin PrepAverage seal resistance is increased when a50:50 trypsin/versene mix is used to lift the cellscompared to that of versene aloneAverage current amplitude is increased when a50:50 AmphoB/Nystatin mix is used to as theperforating agent relative to just AmphoB
Variables influencing assay success• Cell husbandry and preparation– Cell Passage– Cell Density– Trypsin/versene cocktail to lift cells– Slow spin prior to final suspension– Gentle trituration to break-up pellet prior toexperiment start* * * Gentle Gentle Gentle * * *• Compound preparation– Glass vials– Low-binding compound plates– All dilutions made in DMSO
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: Growth kinetics dictate experimenta
Page 13 and 14: The importance of voltage control o
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

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

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?

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