| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 335/460 |
| MW | 331.28 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | NeuroBIOLOGy ReactiveOxygenSpecies |
| Related | CellFunctionalAnalysis RedoxEnzymes |
| Spectrum | AdvancedSpectrumViewer |
1. PrepareThioliteBlue™AMworkingsolution:
a. Preparea1to5mMstocksolutionofThiolite™Blueinhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionneedbealiquotedandfrozenat-20°C.
b. Note:TheunusedThioliteBluestocksolutionshouldbedividedintosingleusealiquotsandstoredat-20oC,protectedfromlight.
2. Prepare1Xdyeworkingsolution
a. Preparea1XdyeworkingsolutionbydilutingtheDMSOstocksolution(fromStep1)inHanksand20mMHepesbuffer(HHBS)orthebufferofyourchoice,pH7rightbeforeuse.Mixthemwellbyvortexing.
b. Note:Thefinalconcentrationofthedyeworkingsolutionshouldbeempiricallydeterminedfordifferentcelltypesand/orexperimentalconditions.Itisrecommendedtotestattheconcentrationsthatareatleastoveratenfoldrange.TherecommendconcentrationinJurkatcellsis1-10uM.
3. Analyzecellswithaflowcytometerorafluorescencemicroscope:
a. Treatcellswithtestcompoundsforadesiredperiodoftime.
b. Centrifugethecellstoget1-5×105cellspertube.
c. ResUSPendcellsin1mLofthedyeworkingsolution(fromStep2).Optional:OnecanaddtheDMSOstocksolutionintothecellsdirectlywithoutmediumremoving(suchas,add1µLof1mMDMSOstocksolutioninto1mLcells)
d. Incubatecellswithadyesolutionatroomtemperatureor37°Cfor10to30min,protectedfromlight.
e. Removethedyeworkingsolutionfromthecells,washthecellswithHHBSorbufferofyourchoice.Resuspendcellsin1mLofpre-warmedHHBSormediumtoget2-10×105cellspertube.
f. MonitorthefluorescencechangeatEx/Em335/460nmwithaflowcytometerorafluorescencemicroscope.
| References&Citations |  CitationExplorer |
GasChromatography/MassSpectrometry-BasedMetabolomicProfilingRevealsAlterationsinMousePlasmaandLiverinResponsetoFavaBeans
Authors:ManXiao,GuankuiDu,GuobingZhong,DongjingYan,HuazongZeng,WangweiCai
Journal:PloSone(2016):e0151103
Virus-likeparticleswithremovablecyclodextrinsenableglutathione-triggereddrugreleaseincells
Authors:KenichiNiikura,NaotoshiSugimura,YusukeMusashi,ShintaroMikuni,YasutakaMatsuo,ShintaroKobayashi,KeitaNagakawa,ShukoTakahara,ChieTakeuchi,HirofumiSawa
Journal:Molecularbiosystems(2013):501--507
