PDD-00017273-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEPDD00017273Cat.No.:HY-108360CASNo.:1945950-21-9分?式:C??H??N?O?S?分?量:514.62作?靶點:Poly(ADP-ribose)Glycohydrolase(PARG)作?通路:CellCycle/DNADamage儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數據體外實驗DMSO:25mg/mL(48.58mM;ultrasonicandwarmingandheatto60°C)MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.9432mL9.7159mL19.4318mL5mM0.3886mL1.9432mL3.8864mL10mM0.1943mL0.9716mL1.9432mL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%salineSolubility:≥2.5mg/mL(4.86mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.86mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.86mM);ClearsolutionBIOLOGICALACTIVITY?物活性PDD00017273?種有效的聚(腺苷?磷酸核糖)?解酶(PARG)抑制劑，IC50值為26nM，KD值為1.45nM。IC50&TargetIC50:26nM(PARG)[1]KD:1.45nM(PARG)[1]體外研究PDD00017273isapotentinhibitorofPARG,withanIC50of26nM,andaKDof1.45nM.PDD00017273(10μM)doesnotinhibitfivecommonCytochromeP450enzymes.PDD00017273(30μM)modestlyincreasessphosphorylatedH2AX(γH2AX)intensity,PDD00017273alsodecreasesinNAD/HthroughPARGinhibitionafterDNAdamage.PDD00017273suppressestheZR-75-1cellscarringBRCA1andBRCA2wildtype,andexhibitslesspotentactivitiesagainstMDA-MB-436cellscarrythe5396+1G>AmutationinBRCA1[1].PDD00017273(0.3μM)inhibitsdegradationofPARpolymersinMCF7cells.PDD00017273(0.3μM)alsoreducestheviabilityofBRCA1,BRCA2,PALB2,FAM175A,andBARD1depletedcells.PDD00017273stallsreplicationforksandinducesDNAdamagethatrequireshomologousrecombination(HR)forrepair[2].PROTOCOLKinaseAssay[1]Briefly,PARGinvitroassaysareconductedinatotalvolumeof15μLinastandard384-wellformat.Atotalof5μLofhumanfulllengthPARGusedatafinalreactionconcentrationof65pM,isaddedto5μLofBt-NADribosylatedPARP1substrateatafinalreactionconcentrationof4.8nMinassaybuffer(50mMTrispH7.4,0.1mg/mLBSA,3mMEDTA,0.4mMEGTA,1mMDTT,0.01%Tween20,50mMKCl).ThereactionisincubatedatRTfor10min,andthen5μLofdetectionreagentisadded.Detectionreagentconsistsof42nMmAbanti-6HISXL665and2.25nMstreptavidineuropiumcryptate,bothat3×workingstockconcentrations(finalconcentrationsof14nMand0.75nM,respectively),indetectionbuffer(50mMTrispH7.4,0.1mg/mLBSAand100mMKF).FollowingincubationatRTfor60mininthedark,TR-FRETsignalismeasuredatλEx340nmandλEm665nmandλEm620nmusingaPHERAstarFSplatereader.Theratioiscalculatedas[Em665/EM620]×104foreachwellandusedtocalculatepercentinhibitionfortestcompounds[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]HeLacellsareseededin30μLofmediaat1×104cells/mLinGreiner384-wellplates.Atotalof16-24hlater,cellsaretreatedwithinhibitors(8ptdoseresponse,0.01-30μM,triplicates)orvehicle(DMSO)control.Theouterwellsareleftundosedtoaccountforedgeeffects.After72h,50μLof3.7%formaldehyde/PBSisaddedtoeachwell,andcellsarefixedfor20min.CellsarethenrinsedtwicewithPBSandstainedfor1hwithHoechst33342/PBS(1:2000)inthedark.AftertwofurtherrinseswithPBS,imagesarecapturedand2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEnucleicountedonaCellInsight.Themaximumnumberoffieldsarecapturedfromeachtriplicatewell,whichapproximatedtoatleast1000nucleiinvehicle-dosedwells[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產品發(fā)表的科研?獻?CellRep.2021Oct5;37(1):109695.?Elife.2022Apr27;11:e72464.?Viruses.2022Sep15;14(9):2049.?CancerRep.26August2022.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].JamesDI,etal.First-in-ClassChemicalProbesagainstPoly(ADP-ribose)Glycohydrolase(PARG)InhibitDNARepairwithDifferentialPharmacologytoAZD2281.ACSChemBiol.2016Nov18;11(11):3179-3190.Epub2016Oct12.[2].GravellsP,etal.SpecifickillingofDNAdamage-responsedeficientcellswithinhibitorsofpoly(ADP-ribose)glycohydrolase.DNARe