Z-VAD-OMe-FMK-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemEZ-VAD(OMe)-FMKCat.No.:HY-16658CASNo.:187389-52-2分⼦式:C₂₂H₃₀FN₃O₇分⼦量:467.49Sequence:Z-Val-Ala-Asp(OMe)-FMKSequenceZVA-D(OMe)-FMKShortening:作⽤靶点:Caspase作⽤通路:Apoptosis储存⽅式:Powder-80°C2years-20°C1yearInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:100mg/mL(213.91mM;Needultrasonic)扫描⼆维码，运⽤溶解⽅案计算器获得适合您实验体系的溶解⽅案MassSolvent1mg5mg10mgConcentration制备储备液1mM2.1391mL10.6954mL21.3908mL5mM0.4278mL2.1391mL4.2782mL10mM0.2139mL1.0695mL2.1391mL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，并请注意储备液的保存⽅式和期限。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：1%DMSO99%saline1/4www.MedChemEwww.MedChemE2.Solubility:≥0.52mg/mL(1.11mM);Clearsolution请依序添加每种溶剂：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.5mg/mL(5.35mM);Clearsolution此⽅案可获得≥2.5mg/mL(5.35mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL25.0mg/mL的澄DMSO储备液加到400μLPEG300中，混合均匀；向上述3.体系中加⼊50μLTween-80，混合均匀；然后继续加⼊450μL⽣理盐⽔定容⾄1mL。请依序添加每种溶剂：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(5.35mM);Clearsolution此⽅案可获得≥2.5mg/mL(5.35mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL25.0mg/mL的澄DMSO储备液加到900μL20%的SBE-β-CD⽣理盐⽔⽔溶4.液中，混合均匀。请依序添加每种溶剂：10%DMSO90%cornoilSolubility:≥2.5mg/mL(5.35mM);Clearsolution此⽅案可获得≥2.5mg/mL(5.35mM，饱和度未知)的澄溶液，此⽅案不适⽤于实验周期在半个⽉以上的实验。5.以1mL⼯作液为例，取100μL25.0mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。请依序添加每种溶剂：5%DMSO40%PEG3005%Tween-8050%saline6.Solubility:≥2.62mg/mL(5.60mM);Clearsolution请依序添加每种溶剂：5%DMSO95%(20%SBE-β-CDinsaline)Solubility:≥2.62mg/mL(5.60mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Z-VAD(OMe)-FMK(Z-Val-Ala-Asp(OMe)-FMK)⼀种不可逆的pan-caspase抑制剂。Z-VAD(OMe)-FMK泛素C末端⽔解酶L1(UCHL1)抑制剂。Z-VAD(OMe)-FMK通过靶向UCHL1活性位点对UCHL1进⾏不可逆地修饰[2]。IC50&TargetCaspase体外研究Z-VAD(OMe)-FMK(Z-Val-Ala-Asp(OMe)-FMK)isabroad-spectrumcaspaseinhibitor,hasbeenshowntoinhibittheintracellularactivationofcaspase-likeproteases.TheinjectionofZ-VAD(OMe)-FMKsuppressesthecaspase-3activityinlungtissues,andsignificantlydecreasesthenumberofterminaldUTPnick-endlabeling-positivecells[1].Z-VAD(OMe)-FMKeffectivelyinhibitsUCHL1'sreactionwithhemagglutinin-taggedubiquitinvinylmethylester(HA-UbVME)attheconcentrationof100μM[2].Z-VAD(OMe)-FMKisadministeredintraperitoneallyat1hourbeforeand6hoursafterSAH.Expressionofcaspase-3andpositiveTUNELisexaminedasmarkersforapoptosis.Z-VAD(OMe)-FMKsuppressesTUNELandcaspase-3staininginendothelialcells,decreasescaspase-3activation,reducesBBBpermeability,relievesvasospasm,abolishesbrainedema,andimprovesneurologicaloutcome[3].Z-VAD(OMe)-FMKisacell-permeablecaspaseinhibitor,efficientlyblockscelldeathinducedbySMNdeficiency[4].体内研究ThesurvivalrateofmiceisprolongedsignificantlybytheinjectionofZ-VAD(OMe)-FMK(Z-Val-Ala-Asp(OMe)-FMK).AllmicesuccumbedtoLPSwithin30hours.Bycontrast,themicetreatedwithZ-VAD(OMe)-FMKsurvivesignificantlylongerand27%ofthemicesurvivedmorethan7days[1].2/4www.MedChemEwww.MedChemEPROTOCOLCellAssay[4]PCRproductscontainingcodingsequencesforthedSMN(forwardprimer:5′-TAATACGACTCACTATAGGGAAGACGTACGACGAGTCG-3′;andreverseprimer:5′-TAATACGACTCACTATAGGGGTGGTGCTGGCTTCTTTC-3′;productlength,601bps;boldanditalicslettersrepresentT7promotersequences)andcontrolDrosophilaPresenilin(dPsn)gene(forwardprimer:5′-TAATACGACTCACTATAGGGTGGCTGCTGTCAATCTC-3′;andreverseprimer:5′-TAATACGACTCACTATAGGGCGATAGCAACGCTTCTTG-3′;productlength:543bps)areobtainedandgel-purified.Double-strandedRNAs(dsRNA)aregeneratedbytranscriptionwithRibomaxT7TranscriptionkitanddigestedwithRnase-freeDNase.ThedsRNAproductsareethanolprecipitatedandannealedbyincubationat65°Cfor30minandthenslowlyallowedtocoolatroomtemperature.TheannealeddsRNAproductsareanalyzedona1%agaorsegeltoensurethemajorityofdsRNAexistedasasingleband.ThedsRNA(2μg)and/orplasmidDNAs(2μg)areintroducedintocellsbyusingCellfectin.Caspaseinhibitionisachievedbyusing50μMofZ-VAD(OMe)-FMKintheculturemedium[4].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1][3]Miceusedinthisstudyare5-to6-week-old(20to22g)ICRmales.Miceareinjectedwith30mg/kgLPSfromE.coliserotypeO111:B4throughthetailvein.AsingleintravenousinjectionofZ-VAD(OMe)-FMK(0.25mg)ismade15minutesbeforeLPSinjection,followedbythreeintravenousinjectionsofZ-VAD(OMe)-FMK(0.1mgeach)perhour.Controlmiceareinjectedwiththesamevolumeof1%DMSOinsterilesaline.Rats[3]MaleSprague-Dawleyratsweighing300to350gareanesthetizedwithα-chloralose(40mg/kgIP)andurethane(400mg/kgIP).Animalsareintubated,andrespirationismaintainedwithasmallanimalrespirator.Rectaltemperatureismaintainedat37±0.5°Cwithaheatingpad.Theleftexternalcarotidarteryisisolatedanda4.0monofilamentnylonsutureisinsertedthroughtheinternalcarotidarterytoperforatethemiddlecerebralartery.SAHisconfirmedatautopsyineachrat.Sham-operatedratsunderwentthesameproceduresexceptthatthesutureiswithdrawnafterresistanceisfelt.Z-VAD(OMe)-FMK(50μMper0.3mL)isinjectedintraperitoneallyat1hourbeforeand6hoursafterSAHinduction.Invehiclegroup,ratsunderwentSAHinductionandaretreatedwiththesamevolumeofvehicle(DMSOdilutedinphysiologicalbuffersolution).Notreatmentisappliedinsham-operatedanimals.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•Nature.2020Apr;580(7803):386-390.•Cell.2020Mar5;180(5):941-955.e20.•Cell.2018Sep6;174(6):1477-1491.e19.•CellMetab.2021Feb2;33(2):424-436.e10.•CellRes.2018Dec;28(12):1171-1185.Seemorecustomervalidationsonwww.MedChemEREFERENCES3/4www.MedChemEwww.MedChemE[1].KawasakiM,etal.Protectionfromlethalapoptosisinlipopolysaccharide-inducedacutelunginjuryinmicebyacaspaseinhibitor.AmJPathol.2000Aug;157(2):597-603.[2].ParkS,etal.Neurovascular