SQ22536-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemESQ22536Cat.No.:HY-100396CASNo.:17318-31-9分⼦式:C₉H₁₁N₅O分⼦量:205.22作⽤靶点:AdenylateCyclase作⽤通路:GPCR/GProtein储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:100mg/mL(487.28mM;Needultrasonic)H2O:55mg/mL(268.01mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM4.8728mL24.3641mL48.7282mL5mM0.9746mL4.8728mL9.7456mL10mM0.4873mL2.4364mL4.8728mL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>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:≥2.5mg/mL(12.18mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(12.18mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(12.18mM);Clearsolution4.请依序添加每种溶剂：PBSSolubility:25mg/mL(121.82mM);Clearsolution;NeedultrasonicBIOLOGICALACTIVITY⽣物活性SQ22536⼀种有效的腺苷酸环化酶(AC)抑制剂。IC50&Targetadenylatecyclase(AC)[1]体外研究SQ22536(SQ22,536)effectivelyinhibitstheeffectofforskolinwithrespectiveIC50valuesof5μM.PreincubationwithgradedconcentrationsofSQ22536revealsthatbothSQ22536effectivelyinhibitsPACAP-inducedreportergeneactivationwithapproximateIC50valueof5μM.SQ22536morepotentlyinhibitsforskolin-inducedElkactivation(IC50=10μM)than8-Br-cAMP-inducedElkactivation(IC50=170μM).Mostnotably,therearesubstantialdifferencesinthereportedpotenciesofSQ22536toinhibittheactivitiesofrecombinantAC5andAC6,withrespectiveIC50valuesof2μMand360μM.Atagreaterconcentration(500μM),SQ22536significantlyinhibitsneuriteelongationduetoeitherforskolinor8-Br-cAMP[1].PROTOCOLCellAssay[1]HEK293CRE-luc2PGloResponseluciferasereportercellsaretransducedwithretroviralvectorsexpressingratPAC1hopreceptors.Individualcelllinesareobtainedbylimitingdilutioncloning,andaclonalPAC1-expressinglineispropagatedandusedforCREluciferaseassays.Inbrief,HEK293CRE-luc2Pcellsareplatedin96-wellplates(10,000cellsin80μLmediaperwell)inassaymedia(DMEMsupplementedwith1%fetalbovineserum).Onedayafterplating,cellsaretreatedwithACinhibitors(10μLinassaymedia/well)for30minutes,followedbyagonists(10μLinassaymedia/well),andareincubatedfor4hours.Luciferaseactivityisdeterminedaftertheadditionof100μL/wellBright-GloLuciferaseAssayReagent.Luminescence(RLU)ismeasuredinaVictor3microtiterplatereaderafter2minutesofagitationatroomtemperature.CyclicAMPismeasuredinNS-1cells.Inbrief,NS-1cellsareseededandgrownovernightin96-wellplates.Thenextday,cellsarepretreatedfor20minutesinmediacontainingthephosphodiesteraseinhibitor3-isobutyl-1-methylxanthine(0.5mM)withorwithoutSQ22536.Afterpretreatmentwithinhibitors,cellsarestimulatedwithagonists,addedas10×solutions,foranadditional20minutes.IntracellularcAMPisthenassayedusingthecAMPBiotrakenzymeimmunoassaykitformeasurementofnonacetylatedcAMP[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•ProgNeurobiol.2021Mar22;102041.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE•CellDeathDis.2020May26;11(5):394.•IntJMolSci.2022,23(20),12595.•IntJMolSci.2022Sep14;23(18):10709.•iScience.2021,102812.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].EmeryAC,etal.AnewsiteandmechanismofactionforthewidelyusedadenylatecyclaseinhibitorSQ22,536.MolPharmacol.2013Jan;83(1):95-105.McePdfHe