Phorbol-12-myristate-13-acetate-PMA-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEPhorbol12-myristate13-acetateCat.No.:HY-18739CASNo.:16561-29-8Synonyms:PMA;TPA;Phorbolmyristateacetate分⼦式:C₃₆H₅₆O₈分⼦量:616.83作⽤靶点:PKC;SphK;NF-κB作⽤通路:Epigenetics;TGF-beta/Smad;Immunology/Inflammation;NF-κB储存⽅式:4°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性数据体外实验DMSO:100mg/mL(162.12mM;Needultrasonic)Ethanol:100mg/mL(162.12mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM1.6212mL8.1060mL16.2119mL5mM0.3242mL1.6212mL3.2424mL10mM0.1621mL0.8106mL1.6212mL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(protectfromlight)。-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)1/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(4.05mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:2.5mg/mL(4.05mM);Suspendedsolution;Needultrasonic3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.05mM);Clearsolution4.请依序添加每种溶剂：10%EtOH>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(4.05mM);Clearsolution5.请依序添加每种溶剂：10%EtOH>>90%(20%SBE-β-CDinsaline)Solubility:2.5mg/mL(4.05mM);Suspendedsolution;Needultrasonic6.请依序添加每种溶剂：10%EtOH>>90%cornoilSolubility:≥2.5mg/mL(4.05mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Phorbol12-myristate13-acetate(PMA)⼀种佛波酯，蛋⽩激酶C(PKC)和SphK的激活剂。Phorbol12-myristate13-acetateNF-κB激活剂。Phorbol12-myristate13-acetate可诱导THP1细胞分化。IC50&TargetPKCNF-κB11.7nM(EC50)体外研究InordertoexaminetheroleofPKCinp38MAPKphosphorylation,thecellsarestimulatedwiththePKCactivator,PMA(100nM),whichmimicsthebindingofDAG,thenaturalactivatorofPKC,totheC1regionofthePKCs.p38MAPKphosphorylationbyPMAisobservedinthetwocelltypessimilartothatobservedbyGnRHinαT3-1cells,thatis,aslowsustainedactivation(3.2-foldand3.6-fold,respectivelyat30min).TheparadoxicalfindingsthatPKCsactivatedbyGnRHandPMAplayadifferentialroleinp38MAPKphosphorylationmaybeexplainedbydifferentiallocalizationofthePKCs.Basal,GnRH-andPMA-stimulationofp38MAPKphosphorylationinαT3-1cellsismediatedbyCa2+influxviavoltage-gatedCa2+channelsandCa2+mobilization,whileinthedifferentiatedLβT2gonadotropecellsitismediatedonlybyCa2+mobilization[2].THP-1cellsaredifferentiatedintomacrophage-likecells(THP-1macrophages)byincubationinthepresenceofPMA(200ng/mL;1-5days),whichleadstoamacrophage-likephenotypecharacterizedbychangesinmorphologyandincreasedcellsurfaceexpressionofCD11andCD14[3].InthemonocyticcelllineTHP-1,PMAresultsinamoredifferentiatedphenotypethanVD3,accordingtoadherence,lossofproliferation,phagocytosisoflatexbeads,andexpressionofCD11bandCD14[5].体内研究PMAisaPKCagonist,whichreversesthedamageinducedby5-hydroxydecanoicacid(5-HD).Thus,activationofthemitoKATPprotectedmitochondrialfunctioninSODandMDAviathePKCpathway[4].PROTOCOLCellAssay[2]αT3-1andLβT-2cellsaregrowninmonolayerculturedinDMEMinhumidifiedincubator5%CO2at37°C.2/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESerumstarvationiswith0.1%FCSinthesamemediumfor16h.GnRHandPMAarethenaddedforthelengthoftimeasindicated.Ingeneral,αT3-1cellsaretransientlytransfectedbyExGen500orbyjetPRIME,whileLβT2cellsonlybyjetPRIMEtransfectionreagent.Forexperimentswithdominant-negative(DN)PKCs,αT3-1cells(in6cmplates)aretransfectedwith1.5μgofp38α-GFPwith3μgofcontrolvector,pCDNA3,orwith3μgoftheDN-PKCsconstructs.ForLβT2cells,transfectionsareperformed(in10cmplates)with4μgofp38α-GFPalongwith9μgofcontrolvector,pCDNA3,orwith9μgoftheDN-PKCsconstructs.Approximately30haftertransfection,thecellsareserumstarved(0.1%FCS)for16handlaterstimulatedwithGnRHorPMA,washedtwicewithice-coldPBS,treatedwiththelysisbuffer,followedbyonefreeze-thawcycle.Cellsareharvested;followingcentrifugation(15,000×g,15min,4°C)supernatantsaretakenforimmunoprecipitationexperiments[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalRats[3]Administration[3]AllexperimentsqreperformedwithmaleWistarrats(weighing250-280g).Onehundredandthirty-fiveWistarratsarerandomlydividedintosevengroups.(1)Ratsintheshamgroup(n=21)aregivenalateralcerebralventricleinjectionof0.9%normalsaline;(2)RatsintheIRgroup(n=21)aregivenalateralcerebralventricleinjectionof0.9%normalsaline30minbeforemiddlecerebralarteryocclusion(MCAO);(3)RatsintheCarbenoxolone(CBX)group(n=21)aregivenalateralcerebralventricleinjectionofCBX(5μg/mL×10μL)30minbeforeMCAO;(4)RatsintheSch-6783group(n=21)aregivenalateralcerebralventricleinjectionofDZX(2mM×30μL)30minpriortoMCAO;(5)Ratsinthe5-HDgroup(n=21)aregivenalateralcerebralventricleinjectionof5-HD(100mM×10μL),andafter10min,DZXisinjected15minpriortoMCAO;(6)TheratsintheDZX+Rogroup(n=15)aregivenalateralcerebralventricleinjectionofDZX,andafter10min,Ro-31-8425(400μg/kg)isinjected15minpriortoMCAO;(7)Theratsinthe5-HD+PMAgroup(n=15)aregivenanintraperitonealinjectionofPMA(200μg/kg)aftertheinjectionof5-HDandDZX.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•MilMedRes.2022Aug23;9(1):46.•JMedVirol.2020Aug10.•EmergMicrobesInfect.2022Oct13;1-49.•MolCell.2022May5;S1097-2765(22)00327-6.•NatCommun.2022Aug15;13(1):4795.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].SergioE.Alvarez,etal.Autocrineandparacrinerolesofsphingosine-1-phosphate.TRENDSinEndocrinologyandMetabolismVol.18No.8[2].MugamiS,etal.DifferentialrolesofPKCisoforms(PKCs)andCa2+inGnRHandphorbol12-myristate13-acetate(PMA)stimulationofp38MAPKphosphorylationinimmortalizedgonadotropecells.MolCellEndocrinol.2017Jan5;439:141-154.[3].HouS,etal.MechanismofMitochondrialConnexin43'sProtectionoftheNeurovascularUnitunderAcuteCerebralIschemia-3/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEReperfusionInjury.IntJMolSci.2016May5;17(5).pii:E679.[4].ZhangT,etal.MPTP-InducedDepletioninBasolateralAmygdalaviaDecreaseofD2RActivationSuppressesGABAAReceptorsExpressionandLTDInductionLeadingtoAnxiety-LikeBehaviors.FrontMolNeurosci.2017Aug