| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 573/588 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | GPCR CalciumGPCRAssays |
| Related | CalciumChannels pHandIonIndicators |
| Spectrum | AdvancedSpectrumViewer |
- Preparecells:
- Foradherentcells:Platecellsovernightingrowthmediumat40,000to80,000cells/well/100µLfora96-wellplateor10,000to20,000cells/well/25µLfora384-wellplate.
- Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletincellgrowthmediumorHHBSat125,000to250,000cells/well/100µLfora96-wellpoly-Dlysineplateor30,000to60,000cells/well/25µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.
Note:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensityfortheintracellularcalciummobilization.
- PrepareCalbryte™590AMdye-loADIngsolution:
- Thawallthekitcomponentsatroomtemperaturebeforeuse.
- MakeCalbryte™590AMstocksolution:Add20µL(forCat#36200)or200µL(forCat#36201and36202)ofDMSOintothevialofCalbryte™590AM(ComponentA),andmixthemwell.
Note:20µLofCalbryte™590AMstocksolutionisenoughforoneplate.UnusedCalbryte™590AMstocksolutioncanbealiquotedandstoredat<-20=""oc=""for=""more=""than=""one=""month=""if=""the=""tubes=""are=""sealed=""tightly.=""protect=""from=""light=""and=""avoid=""repeated=""freeze-thaw=""> - Make1Xassaybuffer:Mix9mLofHHBS(ComponentC,notincludedinthekitCat#36202)with1mLof10XPluronicF127Plus(ComponentB),andmixthemwell.
- MakeCalbryte™590AMdye-loadingsolutionforonecellplate:Add20µLofCalbryte™590AMstocksolution(fromStep2.2)into10mLof1Xassaybuffer(fromStep2.3),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.
- Runcalciumassay:
- Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofCalbryte™590AMdye-loadingsolution(fromStep2.4)intothecellplate.
- Incubatethedye-loadingplateinacellincubatorfor~60minutes,andthenincubatetheplateatroomtemperatureforanother15minutes.
Note1:Iftheassayrequires37oC,performtheexperimentimmediatelywithoutfurtherroomtemperatureincubation.
Note2:Ifthecellscanfunctionwellatroomtemperatureforlongertime,incubatethecellplateatroomtemperaturefor1hour. - PreparethecompoundplatewithHHBSoryourdesiredbuffer.
- RunthecalciumfluxassaybymonitoringthefluorescenceintensitywithTRITCFilter(orEx/Em=540/590nm).
| References&Citations |  CitationExplorer |
CalreticulinregulatesTGF-β1-inducedepithelialmesenchymaltransitionthroughmodulatingSmadsignalingandcalciumsignaling
Authors:YanjiaoWu,XiaoliXu,LunkunMa,QianYi,WeichaoSun,LilingTang
Journal:TheInternationalJournalofBiochemistry&CellBIOLOGy(2017)
Dexmedetomidinereduceshypoxia/reoxygenationinjurybyregulatingmitochondrialfissioninrathippocampalneurons
Authors:JiaLiu,QingDu,HeZhu,YuLi,MaodongLiu,ShoushuiYu,ShileiWang
Journal:IntJClinExpMed(2017):6861--6868
Monosialoganglioside1mayalleviateneurotoxicityinducedbypropofolcombinedwithremifentanilinneuralstemcells
Authors:JiangLu,Xue-qinYao,XinLuo,YuWang,SookjaKimChung,He-xinTang,ChiWaiCheung,Xian-yuWang,ChenMeng,QingLi
Journal:NeuralRegenerationResearch(2017):945
Obtainingspontaneouslybeatingcardiomyocyte-likecellsfromadipose-derivedstromalvascularfractionsculturedonenzyme-crosslinkedgelatinhydrogels
Authors:GangYang,ZhenghuaXiao,XiaomeiRen,HaiyanLong,KunlongMa,HongQian,YingqiangGuo
Journal:ScientificReports(2017):41781
Di(2-ethylhexyl)phthalate-inducedapoptosisinratINS-1cellsisdependentonactivationofendoplasmicreticulumstressandsuppressionofantioxidantprotection
Authors:XiaSun,YiLin,QianshengHuang,JunpengShi,LingQiu,MeiKang,YajieChen,ChaoFang,TingYe,SijunDong
Journal:Journalofcellularandmolecularmedicine(2015):581--594
Theeffectofmitochondrialcalciumuniporteronmitochondrialfissioninhippocampuscellsischemia/reperfusioninjury
Authors:LantaoZhao,ShuhongLi,ShileiWang,NingYu,JiaLiu
Journal:Biochemicalandbiophysicalresearchcommunications(2015):537--542
FungusinducesthereleaseofIL-8inhumancornealepithelialcells,viaDectin-1-mediatedproteinkinaseCpathways.
Authors:Xu-DongPeng,Gui-QiuZhao,JingLin,NanJiang,QiangXu,Cheng-ChengZhu,Jain-QiuQu,LinCong,HuiLi
Journal:Internationaljournalofophthalmology(2014):441--447
Propofolandremifentanilatmoderateandhighconcentrationsaffectproliferationanddifferentiationofneuralstem/Progenitorcells
Authors:QingLi,JiangLu,XianyuWang
Journal:Neuralregenerationresearch(2014):2002
Roleofmitochondrialcalciumuniporterinregulatingmitochondrialfissioninthecerebralcortexesoflivingrats
Authors:NanLiang,PengWang,ShileiWang,ShuhongLi,YuLi,JinyingWang,MinWang
Journal:JournalofNeuralTransmission(2014):593--600
Increasedexpressionofcelladhesionmolecule1bymastcellsasacauseofenhancednerve--mastcellinteractioninahapten-inducedmousemodelofatopicdermatitis
Authors:MHagiyama,TInoue,TFuruno,TIino,SItami,MNakanishi,HAsada,YHosokawa,AIto
Journal:BritishJournalofDermatology(2013):771--778
