Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEHSF1ACat.No.:HY-103000CASNo.:1196723-93-9分?式:C??H??N?O?S?分?量:409.52作?靶點:HSP作?通路:CellCycle/DNADamage;MetabolicEnzyme/Protease儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數據體外實驗DMSO:≥150mg/mL(366.28mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.4419mL12.2094mL24.4188mL5mM0.4884mL2.4419mL4.8838mL10mM0.2442mL1.2209mL2.4419mL請根據產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?內使?，-20°C儲存時，請在1個?內使?。體內實驗請根據您的實驗動物和給藥?式選擇適當的溶解?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：(為保證實驗結果的可靠性，澄的儲備液可以根據儲存條件，適當保存；體內實驗的?作液，建議您現?現配，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沉淀、析出現象，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>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥2.5mg/mL(6.10mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:2.5mg/mL(6.10mM);Suspendedsolution;Needultrasonic3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.10mM);ClearsolutionBIOLOGICALACTIVITY?物活性HSF1A?種可滲透細胞的熱休克轉錄因?1(HSF1)活化劑[1]。HSF1A還充當TRiC/CCT的特異性抑制劑。伴侶蛋?TCP-1環(huán)復合物(TRiC)/含TCP-1的伴侶蛋?(CCT)在毒素易位和/或重折疊中起關鍵作?[4]。IC50&TargetHSF1體外研究HSF1Aprotectscellsfromstress-inducedapoptosis,bindsTRiCsubunitsandinhibitsTRiCactivitywithoutperturbationofATPhydrolysis.GeneticinactivationordepletionoftheTRiCcomplexresultsinhumanHSF1activationandHSF1AinhibitsthedirectinteractionbetweenpurifiedTRiCandHSF1invitro.Moreover,fluorescenceanisotropyexperimentsusingFITCcoupledtoHSF1AdemonstratesthatHSF1A-FITCbindstoapurifiedTcp1subunitofTRiCwithanaffinityofapproximately600nM.ThisisvalidatedqualitativelyviatitrationofpurifiedTcp1intobindingreactionscontaining500nMBiotinorHSF1A-Biotin[1].QuantificationbycountingthenumberofcellcontainingaggregatesasafunctionofthetotalnumberofcellsrevealsthatatHSF1Aconcentrationsaslowas2μM,areducednumberofaggregate-containingcellsareobserved.Thefractionofcellscontainingaggregatescontinuedtodecreaseinadose-dependentmannersuchthatpretreatmentwith12μMHSF1Aresultain~20%ofthecellsexhibitingaggregatesvisiblebyfluorescencemicroscopy[2].體內研究HSF1AenhancesHSF1activity,stabilizesHSF1expressionandminimizesDoxorubicin(DOX)-inducedcardiacdamage.WKYratsarechallengedwithDOX(accumulateddose:30?mg/kgw),andDOXcombinedwithHSF1A(100?mg/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?邊的抑制劑?師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(5?mg/kg)for6consecutiveweeksintraperitonealinjectiontoachieveacumulativedoseof30?mg/kg,whichhasbeenwelldocumentedtoachievecardiotoxicity.ThesmallmolecularHSF1activatorHSF1A(100?mg/kg/day)isinjectedintraperitoneally.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產品發(fā)表的科研?獻?ProcNatlAcadSciUSA.2021Feb16;118(7):e2014457118.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].NeefDW,etal.AdirectregulatoryinteractionbetweenchaperoninTRiCandstress-responsivetranscriptionfactorHSF1.CellRep.2014Nov6;9(3):955-66.[2].NeefDW,etal.Modulationofheatshocktranscriptionfactor1asatherapeutictargetforsmallmoleculeinterventioninneurodegenerativedisease.PLoSBiol.2010Jan1