Real-Thiol-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemERealThiolCat.No.:HY-108715CASNo.:2280796-90-7分?式:C??H??N?O?分?量:411.36作?靶點:Others作?通路:Others儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥101mg/mL(245.53mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.4310mL12.1548mL24.3096mL5mM0.4862mL2.4310mL4.8619mL10mM0.2431mL1.2155mL2.4310mL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?。BIOLOGICALACTIVITY?物活性RealThiol?種于可逆反應的熒光探針，可以定量監(jiān)測活細胞中?胱?肽的實時動態(tài)變化。體外研究RealThiol(RT)showsratiometricfluorescenceresponseswithawidedynamicrangewhenreactingwithGlutathione(GSH).RealThiolanditsGSHadduct(RT-GSH)showsfluorescencemaximaat487and562?1/2MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEnmwithexcitationwavelengthsat405and488?nm,respectively.TheKdforthereactionbetweenRealThiolandGSHis3.7?mM.RealThiolhasmuchimprovedquantumyieldsandphotostability.ThewidedynamicrangeofRealThiolallowsformonitoringoftheGSHlevelchangeswithin1to10?mMinbothdirectionsinlivingcells.Gelpermeationchromatography(GPC)analysisshowesthat10%ofRealThiolreactswiththiolatedproteinswhile90%ofRealThiolreactswithGSH[1].Guidelines(Followingisourrecommendedprotocol.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds)[1].1.RealThiol(RT)isdissolvedinDMSOtoobtaina10mMstocksolutionsandfurtherdilutedbeforeuse.2.Cellsareculturedin10%FBSDMEMcontainingdifferentconcentrationsofButhioninesulfoximine(0.9,1.9,3.9,7.8,15.6,31.3,62.5,125,250and500?μM)for72?handharvestedforanalysis.3.Cellsarethensuspendedinfreshmediumcontaining1?μMofRealThiolprobeassingle-cellsuspensions,andkeepthestainingtimebetween9and10?minforeachsample.4.Analyzesampleonaflowcytometer.PEandpacific-bluechannelswereselectedtomeasurethefluorescenceintensitiesofunreactedandreactedRealThiol,respectively.PROTOCOLKinaseAssay[1]RealThiol(RT)stocksolutionisdilutedwithPBStothedesiredconcentrations.Equalvolumes(typically20to100?μL)ofvariousGlutathione(GSH)solutionswithdifferentconcentrationsaremixedwiththeRealThiolsolution10to15?minbeforemeasurement.Allsamplesarepreparedonone96-well/384-wellplatewith3replicateseach.Theplatereaderissettoreadabsorptionofallsamplesfirst.Fluorescentsignalsarethenrecordedatλex=405nm,λem=485?nm,andλex=488?nm,λem=565?nmwithbottomread.Fortestingfluorescentinterferencebyenvironmentalfactors,theRealThiolstocksolutionisdirectlydilutedwithcorrespondingsolutions(forexample,certainpHbufferorglycerolsolution)andmeasuredusingtheplatereaderwiththesamesettingsstatedabove[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].JiangX,etal.Quantitativereal-timeimagingofglutathione.NatCommun.2017Jul13;8:16087.McePdfHeightCaution:Producthasnotbeenfullyv