HSF1A-DataSheet-生命科学试剂-MedChemExpressVIP

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEHSF1ACat.No.:HY-103000CASNo.:1196723-93-9分⼦式:C₂₁H₁₉N₃O₂S₂分⼦量:409.52作⽤靶点:HSP作⽤通路:CellCycle/DNADamage;MetabolicEnzyme/Protease储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥150mg/mL(366.28mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM2.4419mL12.2094mL24.4188mL5mM0.4884mL2.4419mL4.8838mL10mM0.2442mL1.2209mL2.4419mL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(6.10mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:2.5mg/mL(6.10mM);Suspendedsolution;Needultrasonic3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.10mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性HSF1A⼀种可渗透细胞的热休克转录因⼦1(HSF1)活化剂[1]。HSF1A还充当TRiC/CCT的特异性抑制剂。伴侣蛋⽩TCP-1环复合物(TRiC)/含TCP-1的伴侣蛋⽩(CCT)在毒素易位和/或重折叠中起关键作⽤[4]。IC50&TargetHSF1体外研究HSF1Aprotectscellsfromstress-inducedapoptosis,bindsTRiCsubunitsandinhibitsTRiCactivitywithoutperturbationofATPhydrolysis.GeneticinactivationordepletionoftheTRiCcomplexresultsinhumanHSF1activationandHSF1AinhibitsthedirectinteractionbetweenpurifiedTRiCandHSF1invitro.Moreover,fluorescenceanisotropyexperimentsusingFITCcoupledtoHSF1AdemonstratesthatHSF1A-FITCbindstoapurifiedTcp1subunitofTRiCwithanaffinityofapproximately600nM.ThisisvalidatedqualitativelyviatitrationofpurifiedTcp1intobindingreactionscontaining500nMBiotinorHSF1A-Biotin[1].QuantificationbycountingthenumberofcellcontainingaggregatesasafunctionofthetotalnumberofcellsrevealsthatatHSF1Aconcentrationsaslowas2µM,areducednumberofaggregate-containingcellsareobserved.Thefractionofcellscontainingaggregatescontinuedtodecreaseinadose-dependentmannersuchthatpretreatmentwith12µMHSF1Aresultain~20%ofthecellsexhibitingaggregatesvisiblebyfluorescencemicroscopy[2].体内研究HSF1AenhancesHSF1activity,stabilizesHSF1expressionandminimizesDoxorubicin(DOX)-inducedcardiacdamage.WKYratsarechallengedwithDOX(accumulateddose:30ꢀmg/kgw),andDOXcombinedwithHSF1A(100ꢀmg/kgw/day).SupplementationwithHSF1Asignificantlyelevatescardiacfunctionsbacktothelevelsofthecontrolgroup.HSF1AhasbeenshowntostimulatehumanHSF1nucleartranslocation,elevateproteinchaperoneexpressionandameliorateproteinmisfoldingandcelldeathinaneurodegenerativediseasemodel.TheechocardiographicresultsshowthatHSF1AalsoalleviatesDOX-inducedfailuresincardiacfunction[3].PROTOCOLKinaseAssay[1]Proteinextractsaregeneratedfrommammalian,yeastandE.coliculturesusingbiotin-bindingbuffer(20mMHEPES,5mMMgCl2,1mMEDTA,100mMKCl,0.03%NP-40)supplementedwith1%Trition-X100andproteaseinhibitors.Approximately0.5mgofproteinextractisincubatedwith100μMHSF1A-Biotinfor4hat4°CandHSF1A-BiotinassociatedproteinscapturedbywithNeutrAvidinAgaroseResin.Afterwashinginbiotinbindingbufferproteinsareelutedusing50μLbiotinelutionbuffer(100mMTris,150mMNaCl,0.1mMEDTA,2mMD-biotin),resolvedona4-20%SDS,andimmunoblotted.ForpurifiedTRiCandHsp70analyses,5nMproteinisincubatedinbiotin-bindingbuffer+0.5%TritonX-100with100μMbiotinor100μM2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEHSF1A-Biotinfor4hat4°CandcapturedwithNeutrAvidinResin.ForNiNTApurifiedyeastTcp1,differentconcentrationsofTcp10.5μM,1mM,2mM,3mMand4mMin25mMHepespH7.5,150mMNaClareincubatedwith0.5μMBiotinorHSF1A-Biotinfor4hat4°CandcapturedwithNeutrAvidinResin[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]PC12cellsseededintoa96-wellplate(5×104cells/well)aretreatedwithincreasingconcentrationsofHSF1A(2,4,8and12μM)for15h,atwhichtimehttQ74-GFPexpressionisstimulatedbyincubationinthepresenceof1µg/mLDoxycyclinefor5d.CellviabilityisassessedviatheXTTviabilityassay[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalRats[3]Administration[3]Ten-week-oldWistarKyotorats(WKY)areused.Theratsarehousedataconstanttemperature(22°C)ona12-hlight/darkcyclewithfoodandtapwater.Theanimalsarearrangedintothreegroups:WKYrats(thecontrolgroup),DOXratsandDOXratstreatedwithHSF1A.Eachgroupcontainfiveanimals.TheDOXgroupisinjectedwithDOX(5ꢀmg/kg)for6consecutiveweeksintraperitonealinjectiontoachieveacumulativedoseof30ꢀmg/kg,whichhasbeenwelldocumentedtoachievecardiotoxicity.ThesmallmolecularHSF1activatorHSF1A(100ꢀmg/kg/day)isinjectedintraperitoneally.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•ProcNatlAcadSciUSA.2021Feb16;118(7):e2014457118.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].NeefDW,etal.AdirectregulatoryinteractionbetweenchaperoninTRiCandstress-responsivetranscriptionfactorHSF1.CellRep.2014Nov6;9(3):955-66.[2].NeefDW,etal.Modulationofheatshocktranscriptionfactor1asatherapeutictargetforsmallmoleculeinterventioninneurodegenerativedisease.PLoSBiol.2010Jan1