TMA-DPH-solution-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemETMA-DPH(solution)Cat.No.:HY-DY1057CASNo.:115534-33-3分⼦式:C₂₈H₃₁NO₃S分⼦量:461.62作⽤靶点:FluorescentDye作⽤通路:Others储存⽅式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY⽣物活性TMA-DPH(solution)⼀种疏⽔性荧光膜探针(Ex=355nm;Em=430nm)。TMA-DPH能够锚定在细胞表⾯，定位磷脂双分⼦层的不同区域。通过分析TMA-DPH在质膜及膜的亚结构中的荧光极化数值，可以测定细胞膜的流动性。溶剂及浓度：DMSO：10mM体外研究1.Solutionpreparation1.1PreparationofstocksolutionSolvent:DMSOConcentration:10mM(optimizedaccordingtotheexperiment).1.2PreparationofworkingsolutionDilutewithPBSorserum-freemedium(pH7.4)(optimizedaccordingtotheexperiment).Thecorrespondingstocksolutioncanbedilutedaccordingtotheactualsituation.NotethatifthesolventisDMSO,thecytotoxicityofDMSOmustbeconsidered,andasolventcontrolshouldbeprepared;ifthesolventispurewater,theworkingsolutionneedstobefilteredandsterilizedbeforeaddingcells.Note:Theworkingsolutionshouldbepreparedandusedimmediately.Keepitawayfromlight.2.Cellstaining(suspendedcells)2.1CollectcellsbycentrifugationandwashtwicewithPBSfor5minuteseachtime.2.2TMA-DPH(0.5-5μM)inHanksand20mMHepesbufferpH7.4,andincubateitat37°C,for5-30minutes(optimizedaccordingtotheexperiment).2.3Centrifugeat400gfor3-4minutesanddiscardthesupernatant.2.4AddPBStowashthecellstwice,5minuteseachtime.1/2MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.5WashthecellsandresuspendedintheappropriateHepesbufferpHObserveusingafluorescencemicroscopeorflowcytometer.3.Cellstaining(adherentcells)3.1Culturetheadherentcellsonasterilecoverslip.3.2Removethecoverslipfromtheculturemediumandremovetheexcessculturemedium.3.3AddTMA-DPH(0.5-5μM)inHanksand20mMHepesbufferpH7.4,andincubateitat37°C,for5-30minutes(optimizedaccordingtotheexperiment).3.4WashthecellsandresuspendedintheappropriateHepesbufferpHFluorescencemicroscopydetection(Ex/Em=355/430nm).Note:Ifflowcytometryisrequired,thecellsneedtobedigestedwithtrypsinandresuspendedbeforestaining.REFERENCES[1].IllingerD,etal.ThekineticaspectsofintracellularfluorescencelabelingwithTMA-DPHsupportthematurationmodelforendocytosisinL929cells.JCellBiol.1994May;125(4):783-94.[2].IllingerD,etal.AcomparisonofthefluorescencepropertiesofTMA-DPHasaprobeforplasmamembraneandforendocyticmembrane.BiochimBiophysActa.1995Oct4;1239(1):58-66.[3].BenedettiA,etal.Plasmamembranefluidityinisolatedrathepatocytes:comparativestudyusingDPHandTMA-DPHasfluorescentprobes.JGastroenterolHepatol.1989May-Jun;4(3):221-7.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedical