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AAT Bioquest/Screen Quest™ Calbryte-520 Probenecid-Free and Wash-Free Calcium Assay Kit/36317/1 Plate
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 490/525 |
| 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™520NWdye-loADIngsolution:
- Thawallthekitcomponentsatroomtemperaturebeforeuse.
- MakeCalbryte™520NWstocksolution:Add20µL(forCat.#36317)or200µL(forCat.#36318and#36319)ofDMSOintothevialofCalbryte™520NW(ComponentA),andmixthemwell.
Note:20µLofCalbryte™520NWstocksolutionisenoughforoneplate.UnusedCalbryte™520NWstocksolutioncanbealiquotedandstoredat<-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#36319)with1mLof10XPluronic®F127Plus(ComponentB),andmixthemwell.
- MakeCalbryte™520NWdye-loadingsolutionforonecellplate:Add20µLofCalbryte™520NWstocksolution(fromStep2.2)into10mLof1Xassaybuffer(fromStep2.3),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.
- Runcalciumassay:
- Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofCalbryte™520NWdye-loadingsolution(fromStep2.4)intothecellplate.
- Incubatethedye-loadingplateinacellincubatorfor30minutes,andthenincubatetheplateatroomtemperatureforanother15-30minutes.
Note1:Iftheassayrequires37oC,performtheexperimentimmediatelywithoutfurtherroomtemperatureincubation.
Note2:Ifthecellscanfunctionwellatroomtemperatureforlongertime,incubatethecellplateatroomtemperaturefor1hour(Itisrecommendedthattheincubationtimebenolongerthan2hours.) - PreparethecompoundplatewithHHBSoryourdesiredbuffer.
- RunthecalciumfluxassaybymonitoringthefluorescenceintensityatEx/Em=490/525nm.
| References&Citations |  CitationExplorer |
CalreticulinregulatesTGF-β1-inducedepithelialmesenchymaltransitionthroughmodulatingSmadsignalingandcalciumsignaling
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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
