| Ex/Em(nm) | 420/540/460/580 |
| MW | 830.97 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | NeuroBIOLOGy
ReactiveOxygenSpecies
|
| Related | CellSignaling
BiochemicalAssays
|
DAX-J2™Ratio580/460isanewnitricoxide(NO)sensorrecentlydevelopedbyAATBioquest.ItisacellpermeablereagentthatcanmeasurefreeNOandnitricoxidesynthase(NOS)activityinlivingcellsinaratiometricmodeunderphysiologicalconditions.OnceinsidethecelltheblockinggroupsontheDAX-J2reagentarereleasedtoinducefluorescenceratiochangesatwavelengthsof580and460nmuponNOoxidation.Thefluorescenceintensitiesat580nmand460nmcanbedetectedusingthefiltersetsofCy3®/TRITCandBDHorizon™V450/PacificBlue.Thesetwofiltersetsareequippedwithmostofflowcytometersandfluorescencemicroscope.DAX-J2Ratio580/460hasdistinctadvantagesforNOdetectionthanthepopularDAF-2NOprobe:1).ItdoesnotrequireesteraseactivityforNOdetection.DAF-2requiresintracellularesterasestocleaveitsacetategroupsfordetectingNOactivity.ThisesterasedependenceoftencomplicatestheNOdetectionsinceesterseactivitiesareaffectedbycellhealthandmanyotherfactors.2).DAX-J2productexhibitspH-independentfluorescencewhileDAF-2hasitsfluorescencehighlyaffectedbypH.3).DAX-J2Ratio580/460canbemonitoredinaratiometricmode.Movemouseovergridtodisplaywavelength&intensityvalues.
Wavelength(nm)
1) Preparea5to10mMDAX-J2™stocksolutioninDMSO.Make1to10µMworkingsolutionbydilutingtheDMSOstocksolutionintoHankssolutionwith20mMHepesbuffer(HHBS).
2) Treatcellsasdesired.
3) Incubatethecellswith1-10µM(fromStep#1)DAX-J2™orange(cat#16300),Red(cat#16301) orRatio580/460(cat#16310)for20-60minutesat37ºC.
4) Replacethedye-loADIngsolutionwithHHBSbuffer.
5) Analyzethecellswithaproperfluorescenceinstrumentwithbottomreadmode.
Fluorescentreal-timequantitativemeasurementsofintracellularperoxynitritegenerationandinhibition
Authors:ZhenLuo,QinZhao,JixiangLiu,JinfangLiao,RuoguPeng,YuntingXi,ZhenjunDiwu
Journal:AnalyticalBiochemistry(2017)
InducibleNitricOxideSynthase(iNOS)IsaNovelNegativeRegulatorofHematopoieticStem/ProgenitorCellTrafficking
Authors:MateuszAdamiak,AhmedAbdelbaset-Ismail,JosephBMoore,JZhao,AhmedAbdel-Latif,MarcinWysoczynski,MariuszZRatajczak
Journal:StemCellReviewsandReports(2016):1--12
