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AAT Bioquest/Screen Quest™ 10X cell staining buffer with Phenol Red Plus™/36300/10 Plates
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | None/None |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | GPCR CalciumGPCRAssays |
| Related | CalciumChannels pHandIonIndicators BiochemicalAssays |
1 Thaw10Xcellstainingbufferatroomtemperaturebeforeuse.
Note:ItisOKtouseifthebufferhasprecipitates.
2 Prepare1XScreenQuest™CellStainingBuffer:Add1mLof10XScreenQuestTMcellstainingbufferto9mLofHHBS(1XHank’swith20mMHepesbuffer,pH7.0)orbufferofyourchoice,andmixthemwell.
Note:10mLof1Xstainingbufferisenoughforoneplate.Thebufferisstableatroomtemperature.Itisrecommendedtoaliquotandstoreun-used10Xassaybufferat<-20oC.Protectfromlightandavoidrepeatedfreeze-thawcycles.
3 Prepare2XAssaySolution:Addthecellstainingdyestocksolution(ingeneral,itisaconcentratedDMSOsolution)into1XScreenQuest™CellStainingBuffer(fromStep2)tomakethefinalwellconcentration2Xofthedesiredconcentration.
4 Tothemicroplatewelladd2XAssaySolution(fromStep3)whichisthesamevolumeasthecellculturemedium(e.g.,100uL/well/96-wellor25μL/well/384-well).
5 Incubatethecellsina37oC,5%CO2incubator,orasdesired.
Note:ItispossIBLethatthestainingdyemightinterferewiththe1XScreenQuest™CellStainingBuffer.Inthiscase,it’srecommendedtostainthecellsbyadesiredmethod,andrelacethecellstainingsolutionwiththecellgrowthmediumorHHBS.Thenadd100uL/well/96-well(25μL/well/384-well)of1XScreenQuest™CellStainingBufferintothewell.
6 Observethecellswithafluorescencemicroscopeoraplatereaderasrequired.
| References&Citations |  PrinterFriendlyVersion |
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7. EscobarAL,CifuentesF,VergaraJL.(1995)DetectionofCa(2+)-transientselicitedbyflashphotolysisofDM-nitrophenwithafastcalciumindicator.FEBSLett,364,335.
8. FallonKM,ShacklockPS,TrewavasAJ.(1993)DetectioninVivoofVeryRapidRedLight-InducedCalcium-SensitiveProteinPhosphorylationinEtiolatedWheat(Triticumaestivum)LeafProtoplasts.PlantPhysiol,101,1039.
9. GreimersR,TrebakM,MoutschenM,JacobsN,BoniverJ.(1996)Improvedfour-colorflowcytometrymethodusingfluo-3andtripleimmunofluorescenceforanalysisofintracellularcalciumion([Ca2+]i)fluxesamongmouselymphnodeB-andT-lymphocytesubsets.Cytometry,23,205.
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