ML346-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemEML346Cat.No.:HY-18669CASNo.:100872-83-1分?式:C??H??N?O?分?量:272.26作?靶點(diǎn):HSP作?通路:CellCycle/DNADamage;MetabolicEnzyme/Protease儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:12.5mg/mL(45.91mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM3.6730mL18.3648mL36.7296mL5mM0.7346mL3.6730mL7.3459mL10mM0.3673mL1.8365mL3.6730mL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(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動物和給藥?式選擇適當(dāng)?shù)娜芙?案，配制前請先配制澄的儲備液，再依次添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)天使?；澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；以下溶劑前的百分?指該溶劑在您配制終溶液中的體積占?)：1.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥1.25mg/mL(4.59mM);ClearsolutionBIOLOGICALACTIVITY?物活性ML346Hsp70和HSF-1活性的激活劑，針對Hsp70的EC50為4.6μM。ML346恢復(fù)構(gòu)象疾病模型中的蛋1/3MasterofSmallMolecules—您?邊的抑制劑?師www.MedChemE?質(zhì)折疊，?沒有明顯的細(xì)胞毒性或缺乏特異性。ML346誘導(dǎo)了熱休克反應(yīng)(HSR)的因和蛋?質(zhì)效應(yīng)?的特異性增加，包括伴侶蛋?如Hsp70，Hsp40和Hsp27。IC50&TargetHSP704.6μM(EC50,HeLacells)體外研究ML346isanactivatorofHsp70,withanEC50of4600nMinHeLacells.ML346(10μM)restoresproteostasis,restoresCFTR-mediatediodideconductance,andenhancesthecorrectfoldingofproteinsexpressedintwodifferentcellularcompartments[1].ML346(CompoundF1)inducesmultipleresponsesandstronglyinducesHsp70,theoxidativestressresponsegenes(HO1andGCLM),anda2.5-foldupregulationofBiPinWTMEFcells.ML346(0.5-25μM)exhibitscytoprotectiveeffectsincellsaftera35minsevereheatshock,andalsocausesatwo-foldprotectionfromH2O2-inducedapoptosis[2].體內(nèi)研究ML346suppresstheaggregationofpolyQ35inaC.elegansmodel,suggestingtheprobehasefficacyinmodifyingproteinaggregationandassociatedtoxicity[1].PROTOCOLKinaseAssay[2]Inbrief,HeLacellsareincubatedwitheitherDMSO(negativecontrol),thepositivecontrolsMG132(10μM)andlactacystin(6μM)orthePRsA1,A3andML346(F1)for3and6hoursandthenharvested.Cellsarelysedinhomogenizationbuffer(50mMTris-HCl,pH7.5,250mMsucrose,5mMMgCl2,2mMATP,1mMDTT,0.5mMEDTA,0.025%digitonin)for5minonice,andtotalproteinconcentrationofwholecellextractisdetermined.3μgofwholecellextractsarecombinedwithassaybuffer(50mMTris-HCl,pH7.5,40mMKCl,5mMMgCl2,0.5mMATP,1mMDTT,0.05mg/mLBSA)inablack96-wellplateandthereactionisinitiatedbytheadditionofa2×(200μM)fluorogenicpeptidesubstrateSuc-LLVY-AMC.Fluorescenceismeasuredevery10minusingaSynergyH4multi-modemicroplatereader[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]HeLacellsareplatedatadensityof10,000cellsperwellinblack96-wellplatesin100μLofDMEMsupplementedwith10%FBSand1%Pen/Strep/Neo.Platesareincubatedfor16hoursat37°C,5%CO2and95%relativehumiditybeforecompoundaddition.1μLofhitcompounds(ML346)inDMSOorDMSOaloneareaddedtothesampleorcontrolwells,respectively.Platesarethenplacedbackintheincubatorfor24hours.Afterincubation,cellsarewashed2×with200μLofPBSand200μLofasolutionof1μg/mLofcalceinAMisaddedtoeachwell.Cellsarethenincubatedfor45minat37°C,5%CO2beforefluorescencemeasurementusinganAnalystGTmultimodereader.PercentcytotoxicityisexpressedrelativetowellscontainingcellstreatedwithDMSOonly(100%)[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].CalaminiB,etal.ML346:ANovelModulatorofProteostasisforProteinConformationalDiseases.ProbeReportsfromtheNIHMolecularLibrariesProgram.Bethesda(MD):NationalCenterforBiotechnologyInformation(US);2010-.2012Dec17.[2].CalaminiB,etal.Small-moleculeproteostasisregulatorsforproteinconformationaldiseases.NatChemBiol.2011Dec25;8(2):185-96.2/3MasterofSmallMolecules—您?邊的抑制劑?師www.MedChemEMcePd