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TheEchelonAutotaxinInhibitorAssayServiceusesoneofEchelon’sAutotaxinInhibitorScreeningkitsdependingonthescaleoftheproject(catalog#’sK-4200orK-4200HTS).ThiskitusesthefluorogenicATXsubstrate,FS-3.FS-3isanLPCanaloguethatisconjugatedwithbothafluorophore(fluorescein)andaquencher.Initsnativestatethequencherinterfereswithfluorescein’sfluorescence.OnceATXcleavesFS-3,fluoresceinbecomesliberatedfromthequencher,resultinginincreasedfluorescence.
Inthisassayservice,twoscreensarerunforeachsample.Intheinitialscreen,recombinantAutotaxinisincubatedwithyoursampleinhibitor(s)andaninternalcontrolinhibitor(BrP-LPA).Thesubstrate,FS-3,isthenaddedsubsequentlyandfluorescenceismeasuredovertime.Theseassaysarecontinuous(notendpoint)andhavetheadvantageofbeinglesssusceptIBLetoerrorarisingfrommistimingintheadditionofreagents.Thesecondaryscreenisperformedtoevaluatewhetherasamplehit,revealedintheinitialscreen,isafalsepositiveduetointerferencewiththeassay.ThesampleinhibitorsthatarepositiveintheinitialscreenandnegativeinthesecondaryscreenarepotentialAutotaxininhibitors.
Testresultswillbeprovidedas%inhibitionorIC50.Alldatapointswillberuninduplicateandreportedasanaverage.FormoreinformationontheAutotaxinInhibitorScreeningkit,pleaseseeTechnicalDataSheet(TDS)forcatalog#K-4200orK-4200HTS.
SampleSolvent-Theassayissensitivetoorganicsolvents.Limitto<2%.-TheassayisresistanttomodestconcentrationsofDMSO.Seefigure2onK-4200TDS.-Wewillruncontrolsforallsolventsusedinyoursamplepanel.Thismayaddadditionalcosts.-Itmaybepreferredtosendusvialsofyoursamplesolvent.
SampleAmount:60uL/sampleminimumisrequired.Samplesneedtobeinsolutionattheconcentrationstobetested.
| Categories | Services |
|---|---|
| Filter | Autotaxin,Inhibitor |
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