ML335-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEML335Cat.No.:HY-104005CASNo.:825658-06-8分⼦式:C₁₅H₁₄Cl₂N₂O₃S分⼦量:373.25作⽤靶点:PotassiumChannel作⽤通路:MembraneTransporter/IonChannel储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:155mg/mL(415.27mM;Needultrasonicandwarming)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.6792mL13.3958mL26.7917mL5mM0.5358mL2.6792mL5.3583mL10mM0.2679mL1.3396mL2.6792mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.70mM);Clearsolution1/2MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEBIOLOGICALACTIVITY⽣物活性ML335TREK-1和TREK-2的选择性激活剂。IC50&TargetTREK-1,TREK-2体外研究Xenopusoocytetwo-electrodevoltage-clampmeasurementsshowthatML335andML402activateK2P2.1andK2P10.1butnotK2P4.1(14.3±2.7ꢀμM,K2P2.1-ML335;13.7±7.0ꢀμM,K2P2.1-ML402;5.2±0.5ꢀμM,K2P10.1-ML335;and5.9±1.6ꢀμM,K2P10.1-ML402).SwappingtheLys271equivalentbetweenK2P2.1andK2P4.1resultsinaclearphenotypereversalforML335andM402activation.ML335andML402activateK2P2.1inHEK293cellssimilartotheireffectsinXenopusoocytes(5.2±0.8ꢀμMand5.9±1.6ꢀμMforML335andML402,respectively(n≥3))[1].PROTOCOLCellAssay[1]MouseK2P2.1,humanK2P4.1,andmutantsareexpressedfromapreviouslydescribedpIRES2-EGFPvectorinHEK293Tcells(ATTC).70%confluentcellsaretransfected(in35-mmdiameterwells)withLipofectAMINE2000for6ꢀh,andplatedontocoverslipscoatedwithMatrigel.EffectsofML335,ML402andarachidonicacidonK2P2.1currentat0ꢀmVaremeasuredbywhole-cellpatch-clampexperiments24ꢀhaftertransfection.AcquisitionandanalysisareperformedusingpCLAMP9andanAxopatch200Bamplifier.Pipetteresistancerangesfrom1to1.5ꢀMΩ.Pipettesolutioncontainsthefollowing:145ꢀmMKCl,3ꢀmMMgCl2,5ꢀmMEGTAand20ꢀmMHEPES(pH7.2withKOH).Bathsolutioncontainsthefollowing:145ꢀmMNaCl,5ꢀmMKCl,1ꢀmMCaCl2,3ꢀmMMgCl2and20ꢀmMHEPES(pH7.4withNaOH).K2P2.1currentsareelicitedbya1ꢀsrampfrom-100to+50ꢀmVfroma-80ꢀmVholdingpotential.Afterstabilizationofthebasalcurrent,ML335andML402areperfusedat200ꢀmLperhouruntilpotentiationisstablyreached[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].LolicatoM,etal.K2P2.1(TREK-1)-activatorcomplexesrevealacrypticselectivityfilterbindingsite.Nature.2017Jul20;547(7663):364-368.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedica