Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEML335Cat.No.:HY-104005CASNo.:825658-06-8分?式:C??H??Cl?N?O?S分?量:373.25作?靶點:PotassiumChannel作?通路:MembraneTransporter/IonChannel儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:155mg/mL(415.27mM;Needultrasonicandwarming)MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.6792mL13.3958mL26.7917mL5mM0.5358mL2.6792mL5.3583mL10mM0.2679mL1.3396mL2.6792mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；以下溶劑前顯?的百分?指該溶劑在您配制終溶液中的體積占?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.70mM);Clearsolution1/2MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEBIOLOGICALACTIVITY?物活性ML335TREK-1和TREK-2的選擇性激活劑。IC50&TargetTREK-1,TREK-2體外研究Xenopusoocytetwo-electrodevoltage-clampmeasurementsshowthatML335andML402activateK2P2.1andK2P10.1butnotK2P4.1(14.3±2.7?μM,K2P2.1-ML335;13.7±7.0?μM,K2P2.1-ML402;5.2±0.5?μM,K2P10.1-ML335;and5.9±1.6?μM,K2P10.1-ML402).SwappingtheLys271equivalentbetweenK2P2.1andK2P4.1resultsinaclearphenotypereversalforML335andM402activation.ML335andML402activateK2P2.1inHEK293cellssimilartotheireffectsinXenopusoocytes(5.2±0.8?μMand5.9±1.6?μMforML335andML402,respectively(n≥3))[1].PROTOCOLCellAssay[1]MouseK2P2.1,humanK2P4.1,andmutantsareexpressedfromapreviouslydescribedpIRES2-EGFPvectorinHEK293Tcells(ATTC).70%confluentcellsaretransfected(in35-mmdiameterwells)withLipofectAMINE2000for6?h,andplatedontocoverslipscoatedwithMatrigel.EffectsofML335,ML402andarachidonicacidonK2P2.1currentat0?mVaremeasuredbywhole-cellpatch-clampexperiments24?haftertransfection.AcquisitionandanalysisareperformedusingpCLAMP9andanAxopatch200Bamplifier.Pipetteresistancerangesfrom1to1.5?MΩ.Pipettesolutioncontainsthefollowing:145?mMKCl,3?mMMgCl2,5?mMEGTAand20?mMHEPES(pH7.2withKOH).Bathsolutioncontainsthefollowing:145?mMNaCl,5?mMKCl,1?mMCaCl2,3?mMMgCl2and20?mMHEPES(pH7.4withNaOH).K2P2.1currentsareelicitedbya1?srampfrom-100to+50?mVfroma-80?mVholdingpotential.Afterstabilizationofthebasalcurrent,ML335andML402areperfusedat200?mLperhouruntilpotentiationisstablyreached[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].LolicatoM,etal.K2P2.1(TREK-1)-activatorcomplexesrevealacrypticselectivityfilterbindingsite.Nature.2017Jul20;547(7663):364-368.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedica