Z-DEVD-FMK-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemEZ-DEVD-FMKCat.No.:HY-12466CASNo.:210344-95-9分?式:C??H??FN?O??分?量:668.66作?靶點(diǎn):Caspase作?通路:Apoptosis儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥33.33mg/mL(49.85mM)掃描?維碼，*"≥"meanssoluble,butsaturationunknown.運(yùn)?溶解?案計算器獲得適合您實驗體系的溶解?案MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.4955mL7.4776mL14.9553mL5mM0.2991mL1.4955mL2.9911mL10mM0.1496mL0.7478mL1.4955mL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，并請注意儲備液的保存?式和期限。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當(dāng)?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(dāng)保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)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%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.5mg/mL(3.74mM);Clearsolution此?案可獲得≥2.5mg/mL(3.74mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到400μLPEG300中，混合均勻；向上述體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。1/4www.MedChemEwww.MedChemE2.請依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(3.74mM);Clearsolution此?案可獲得≥2.5mg/mL(3.74mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到900μL20%的SBE-β-CD?理鹽??溶3.液中，混合均勻。請依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.5mg/mL(3.74mM);Clearsolution此?案可獲得≥2.5mg/mL(3.74mM，飽和度未知)的澄溶液，此?案不適?于實驗周期在半個?以上的實驗。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到900μL??油中，混合均勻。BIOLOGICALACTIVITY?物活性Z-DEVD-FMK?種特異性的不可逆的caspase-3抑制劑，IC50為18μM[1]。IC50&TargetCaspase-318μM(IC50)體外研究N27cellsareexposedtoMPP+intheabsenceorpresenceof50μMZ-DIPD-FMKor100μMZ-DEVD-FMKor50μMZ-LEHD-FMKandthencaspase-9andcaspase-3enzymaticactivitiesaredeterminedbyenzymaticassayat12and24hfollowingexposure,respectively.Exposureto300μMMPP+for24hinN27cellsresultsinanapproximately2.5-foldincreaseincaspase-3enzymeactivity.MPP+-inducedincreasesincaspase-3enzymeactivityaresignificantlyblockedby50μMZ-DIPD-FMK,100μMZ-DEVD-FMK,and50μMZ-LEHD-FMK[1].體內(nèi)研究EarlyZ-DEVD-FMK(160ng)treatmentimprovesmotorandcognitivefunctionaftertraumaticCNSinjuryinducedbyseverecontrolledcorticalimpact(CCI)inthemouse[2].TreatmentwithZ-DEVD-FMK(160ng)significantlyimprovesneurologicaloutcomewhencomparedwithtraumatizedanimalstreatedwithDMSOvehicle(p<0.01)[3].PROTOCOLCellAssay[1]N27cellsandprimarymesencephalicneuronsareexposedtoeither10-100μM6-OHDAor10-300μMMPP+inthepresenceorabsenceof0.1-50μMZ-DIPD-FMKor0.1-100μMZ-DEVD-FMKor50μMZ-IETD-FMKorZ-LEHD-FMKforthedurationoftheexperiment.N27cellsareincubatedwith100μM6-OHDAfor24hor300μMMPP+for36hinthepresenceorabsenceof50μMZ-DEVD-FMKandcelldeathisdeterminedbyMTTassay,whichiswidelyusedtoassesscellviability.Aftertreatment,thecellsareincubatedinserum-freemediumcontaining0.25mg/mLMTTfor3hat37°C.Formationofformazanfromtetrazoliumismeasuredat570nmwithareferencewavelengthat630nmusingaSpectraMaxmicroplatereader[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[2]Administration[2][3]MaleC57Bl/6mice(20-25g)areused.FortreatmentwithZ-DEVD-fmkorvehicleafterCCI,miceareplaced2/4www.MedChemEwww.MedChemEinastereotaxicapparatus,andtheCCIwoundisreopenedforintracerebroventricularinjection.EitherZ-DEVD-FMK(160ngin2μLDMSO),orDMSOvehicleisinjectedovera5-minuteperiod.Rats[3]MaleSpragueDawleyrats(425±25g)areused.DMSO(5μL)vehicleorZ-DEVD-FMK(160ngin5μLofDMSO)isadministeredatacontrolledrateof0.5μL/minviaaninfusionpumpat30minbeforeandat6and24hrafterTBI.Atthedesignatedtimeperiodsafterinjury,animalsaredecapitatedunderNSC10816anesthesia(100mg/kg,i.p.),andthebrainsareremovedrapidlyanddissected.Sham-operated(control)animalsreceivedanesthesiaandsurgerybutarenotsubjectedtotrauma.Tissuesamplesarecollected1,4,12,24,and72hrafterTBI.Samplesarefrozenondryiceandkeptat?85°C.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?ProcNatlAcadSciUSA.2020Sep22;117(38):23869-23878.?CellMolImmunol.2021Jan;18(1):219-229.?CellRep.2020Nov10;33(6):108369.?ActaPharmSinB.2020Aug;10(8):1397-1413.?Elife.2020Jun2;9:e54954.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].KanthasamyAG,etal.Anovelpeptideinhibitortargetedtocaspase-3cleavagesiteofaproapoptotickinaseproteinkinaseCdelta(PKCdelta)protectsagainstdopaminergicneuronaldegenerationinParkinson'sdiseasemodels.FreeRadicBiolMed.2006Nov15;41(10):1578-89.[2].KnoblachSM,etal.Caspaseinhibitorz-DEVD-fmkattenuatescalpainandnecroticcelldeathinvitroandaftertraumaticbraininjury.JCerebBloodFlowMetab.2004Oct;24(10):1119-32.[3].YakovlevAG,etal.ActivationofCPP32-likecaspasescontributestoneuronalapoptosisandneurologicaldysfunctionaftertraumaticbraininjury.JNeurosci.1997,17(19),7415-7424.[4].HuangMY,etal.ChemotherapeuticagentCPT-11eliminatesperitonealresidentmacrophagesbyinducingapoptosis.Apoptosis.2016Feb;21(2):130-42.McePdfHeight3/4www.MedChemEww