Vari-Fluor-488-Phalloidin-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEVariFluor488-PhalloidinCat.No.:HY-D1817作⽤靶点:FluorescentDye作⽤通路:Others储存⽅式:-20°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)BIOLOGICALACTIVITY⽣物活性VariFluor488-PhalloidinPhalloidin的荧光衍⽣物，可对肌丝蛋⽩特异性标记，在标记时可呈现绿⾊荧光，可使⽤FITC通道进⾏荧光成像(Ex/Em=488nm/513nm)。体外研究GeneralIntrodctionGhostpencyclicpeptideisatoxinisolatedfromthedeadlyumbrella-shapedmushrooms.ItisabicyclicpeptidethatbindsspecificallytoF-actin.Therefore,thedistributionofF-actincanbeeasilystudiedbylabelingthepeptidewithafluorescentdye.Insidethecyclicpeptide,anuncommonthioetherbridgebetweencysteineandtryptophanformsaninnerringstructure.AtelevatedpH,thethioetheriscleavedandthepeptidelosesitsaffinityforactin.VariFluor-PhalloidinisafluorescentderivativeoftheVFseriesofdye-labeledphalloidinthatspecificallylabelsactin.Thefluorescenceishighlystableandcanberetainedinthecellformorethanaweek.Oneunit(T)ofVF-Phalloidinreferstotheamountofdyeusedtostainaslideloadedwithcells.GeneralProtocol1.PreparationofVF-PhalloidinWorkingSolution1.1PreparationofstoragesolutionTakeappropriateamountofmethanolorsterilewatertodissolvethelyophilizedpowderinthebrowntubetoconfigurethemotherliquor.Add0.25mLofdissolutionsolutionto50Tdye.Add1.5mLofsolventto300Tsizedye.Note:VF-Phalloidinstoragesolutionisrecommendedtobestoredat-20℃or-80℃afterdispensingandprotectedfromlight,andmethanolisrecommendedforlong-termstorage.1.2Preparationofworkingsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEDilutethestocksolutionwithpre-warmedserum-freecellculturemediumorPBSat1:40-1:200,i.e.,dilute1-5μLofVF-Phalloidinstocksolutionper200μLofPBS.Note:PleaseadjusttheconcentrationofVF-Phalloidinworkingsolutionaccordingtotheactualsituationanduseitasitis.2.Cellstaining(adherentcells)2.1Cultureadherentcellsonsterilecoverslipsandwashthem3timeswithPBS.2.2FixthecellswithPBSsolutioncontaining4%formaldehydefor20minatroomtemperature.Note:Methanolcandestroyactinduringfixation.Therefore,itisbesttoavoidfixativescontaininganymethanol.Thepreferredfixativeismethanol-freeformaldehyde.2.3Washthecells3timeswithPBS.2.4Thecellsarepermeabilizedwith0.4%TritonX-100inPBSfor10minatroomtemperature.2.5WashthecellsthreetimeswithPBS.2.6Add200μLofdyeworkingsolution,shakegentlytocompletelycoverthecells,andincubateatroomtemperatureawayfromlightfor15-45minutes.Note:Thechromosomevolumecanbeadjustedaccordingtothesample.Toavoidevaporationofthedyesolutionduringincubation,placethecoverslipinasealedcontainer.2.7Pipetteoffthedyeworkingsolutionandwash2-3timeswithPBSorculturemedium.2.8Usefluorescencemicroscopeorflowcytometertoobserveunderthecorrespondingchannel.Precautions1.Fluorescently-labeledGhostPenCyclicPeptidesarenotcellpermeableandthereforehavenotbeenwidelyusedforlivecelllabeling.However,ithasbeenreportedthatlivingcellsmaybelabeledbycytosolicdrinkingorbyunknownmechanisms.Generally,moredyeisrequiredtostainlivingcells.Alternatively,fluorescentlylabeledghostpencyclicpeptidescanbeinjectedintocellstomonitoractindistributionandcellmovement.2.Theactualcontentofthedyeissmall,sodissolveitdirectlyinthetubeuponreceiptandperformtheexperiment.3.Pleasecentrifugetheproductinstantlytothebottomofthetubebeforeuse.4.Thisproductislimitedtoscientificresearchusebyprofessionals,andisnottobeusedforclinicaldiagnosisortreatment,foodormedicine.5.Foryoursafetyandhealth,pleasewearlabcoatanddisposablegloves.户使⽤本产品发表的科研⽂献•MaterTodayBio.2025Mar29.•Phytomedicine.2025May4:143:156830.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].BorovikovYuS,etal.TheeffectofphallotoxinsonthestructureofF-actininmyosin-freeghostmusclefibresofrabbit.FEBSLett.1984Oct29;176(2):441-3.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.Med