UNC3866-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEUNC3866Cat.No.:HY-100832CASNo.:1872382-47-2分?式:C??H??N?O?分?量:795.02作?靶點:HistoneMethyltransferase作?通路:Epigenetics儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥27mg/mL(33.96mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.2578mL6.2891mL12.5783mL5mM0.2516mL1.2578mL2.5157mL10mM0.1258mL0.6289mL1.2578mL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；?旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)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(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當(dāng)?shù)娜芙?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(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)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(xiàn)沉淀、析出現(xiàn)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(3.14mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(3.14mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(3.14mM);ClearsolutionBIOLOGICALACTIVITY?物活性UNC3866?種有效的CBX7-H3拮抗劑，IC50為66±1.2nM。IC50&TargetIC50:66±1.2nM(CBX7)[1]體外研究UNC3866,apotentantagonistofthemethyl-lysine(Kme)readingfunctionofthePolycombCBXandCDYfamiliesofchromodomains.UNC3866bindsthechromodomainsofCBX4andCBX7mostpotentlywithaKdof100nMforeach,andis6-to18-foldselectiveversussevenotherCBXandCDYchromodomainswhilebeinghighlyselectiveversus>250otherproteintargets.UNC3866inhibitsPC3cellproliferation,aknownCBX7phenotype,whileUNC4219,amethylatednegativecontrolcompound,hasnegligibleeffects.UNC3866isapotentandcellularlyactiveantagonistofPRC1chromodomains.UNC3866isthemostpotentligandreportedforCBX7withaKdof97±2.4nM.UNC3866isequipotentforCBX4,whichismostsimilartoCBX7,whileitis18-,6-and12-foldselectiveforCBX4/7overCBX2,-6and-8,respectively.Additionally,UNC3866is65-foldselectiveforCBX4/7overCDY1and9-foldselectiveforCBX4/7overCDYL1bandCDYL2[1].PROTOCOLKinaseAssay[1]TheeffectofUNC3866onthemethyltransferaseactivityofG9a,EHMT1,SUV39H1,SUV39H2,SETDB1,SETD8,SUV420H1,SUV420H2,SETD7,MLL1trimericcomplex,MLL3tetramericcomplex,EZH2trimericcomplex,PRMT1,PRMT3,PRMT5-MEP50complex,PRMT6,PRMT7,PRMT8,PRDM9,SETD2,SMYD2,SMYD3,BCDIN3DandDNMT1isassessedbymonitoringtheincorporationoftritium-labeledmethylgrouptolysineorarginineresiduesofpeptidesubstratesusingScintillationProximityAssay(SPA).Assaysareperformedina20μLreactionmixturecontaining3H-SAMatsubstrateconcentrationsclosetotheKmvaluesforeachenzyme.Threeconcentrations(1μM,10μM,and50μM)ofUNC3866areusedinallselectivityassays.Tostoptheenzymaticreactions,7.5MGuanidinehydrochlorideisadded,followedby180μLofbuffer(20mMTris,pH8.0).Thereactionsaremixedandthentransferredtoa96-wellFlashPlate.ThereactionmixturesinFlashplatesareincubatedfor1hourandtheCPMaremeasuredusingaTopCountplatereader.TheCPMcountsintheabsenceofcompoundforeachdatasetaredefinedas100%activity.Intheabsenceoftheenzyme,theCPMcountsineachdatasetaredefinedasbackground(0%)[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]PC3cellsareseededat200cells/wellinto24-wellplates.Cellsareallowedtoadhereovernight.Themedia(DMEMsupplementedwith10%FBS)isthenexchangedwithfreshmediacontainingDMSO,UNC3866orUNC4219.Ondaythree,themediaareexchangedwithfreshmediacontainingDMSO,UNC3866or2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEUNC4219.Fordose-responsestudies,theEC50isderivedfromasix-pointtitrationrangingfrom100μMto0.4μMofUNC3866orUNC4219.Atday0,3or6,cellsarefixedwithice-coldmethanolfor30sec.andrehydratedwithPBS.NucleiofthecellsarestainedwithDAPI(0.05μg/mL)andnumeratedusingHighContentMicroscopy.Fordose-responsestudies,thecellcountofUNC3866-orUNC4219-treatedcellsisnormalizedtotheaveragecellcountofDMSO-treatedcells.TheEC50iscalculatedusingthe“l(fā)og[inhibitor]vs.thenormalizedresponse-Variableslope”equationinGraphPadPrism5[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?ActaPharmacolSin.2021Apr13.?bioRxiv.2021May26.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].StuckeyJI,etal.AcellularchemicalprobetargetingthechromodomainsofPolycombrepressivecomplex1.N