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AAT Bioquest/Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*/22801/
Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 510/525 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | CellAnalysis CellCytotoxicity |
Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1.Preparecells:
Preparecellsatthedensityfrom5×105to1×106cells/mL.
Note:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensityforapoptosisinduction.
2.PrepareJC-10dye-loADIngsolution:
2.1 Thawallthekitcomponentsatroomtemperaturebeforeuse.
2.2 Add25µLof200XJC-10(ComponentA)into5mLofAssayBuffer(ComponentB),andmixwell.
Note:Aliquotandstoretheunused200XJC-10(ComponentA)at-20oC.Avoidrepeatedfreeze/thawcycles.
3.RunJC-10assay:
3.1 Treatcellswithtestcompoundsforadesiredperiodoftimetoinduceapoptosis.Setupparallelcontrolexperiments.
ForNegativeControl:Treatcellswithvehicleonly.
ForPositiveControl:TreatcellswithFCCPorCCCPat2-10µMina37oC,5%CO2incubatorfor15to30minutes.
Note:CCCPorFCCPcanbeaddedsimultaneouslywithJC-10dye-loadingsolution(fromStep2.2).TitrationoftheCCCPorFCCPmayberequiredforoptimalresultswithanindividualcelllines.
3.2 Centrifugethecellstoget2-5×105cellspertube.
Note:Foradherentcells,gentlyliftthecellsby0.5mMEDTAtoremainthecellsintake,andwashthecellsoncewithserum-containingmediapriortoincubationwithJC-10dye-loadingsolution(seeStep3.3).
3.3 ResUSPendcellsin500μLofJC-10dye-loadingsolution(fromStep2.2).
3.4 Incubatethedye-loadingcellsatroomtemperatureorina37oC,5%CO2incubatorfor15-60minutes,protectedfromlight.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
3.5 MonitorthefluorescenceintensitybyusingaflowcytometryinFL1channelforthegreenfluorescentmonomericsignal(inapoptoticcells),andtheFL2channelfortheorangefluorescentaggregatedsignal(inhealthycells).Gateonthecells,excludingdebris.ItisrecommendedthatcompensationcorrectionsbeperformedusingtheFCCPorCCCP-treatedcells(SeeAppendix).
References&Citations | CitationExplorer |
Below,youmayfindasmallsamplingofspecificJC-10™applications.ToinquireaboutapotentialapplicationofJC-10™,ortoconsultwithourfluorescentdyespecialists,pleasecontactusatsupport@AATbio.comor1-800-990-8053.
- High-contentassaysforhepatotoxicityusinginducedpluripotentstemcell–derivedcells.
ResearchersuseJC-10™tomonitormitochondrialdepolarizationasanindicationofhepatotoxicityandoxidativestressininducedpluripotentstemcell-derivedcells,withtheultimategoalofdesigningareliable,high-contentandimaging-basedinvitrotoxicityassay. - High-ContentHigh-ThroughputAssaysforCharacterizingtheViabilityandMorphologyofHumaniPSC-DerivedNeuronalCultures
JC-10™wasusedtostudyneuronsderivedfrominducedpluripotentstem-cells.SinceJC-10™willaccumulateinthemitochondriaofviablecells,itwasusedtodeterminecellviabilityinahigh-throughputassaycontext. - AnticancerActivityofNewSyntheticα-Methylene-δ-LactonesonTwoBreastCancerCellLines
ResearcherschoseJC-10™toinvestigatemitochondrialmembranepotentialandmembraneintegrityincellstreatedwithnaturalproducts,suchasα-Methylene-δ-Lactones,withthegoalofdevelopingnewtreatmentsforbreastcancer. - Tetrandrineprotectsmouseretinalganglioncellsfromischemicinjury.
JC-10™wasusedinflowcytometrytostudymitochondrialmembranepotential(ΔΨm)inprimaryculturedretinalganglioncells,asanextensionofthefieldofdrugdiscoveryintopreventionofischemicinjury. - Midazolaminducescellularapoptosisinhumancancercellsandinhibitstumorgrowthinxenograftmice
Inastudyofhumancancercells,JC-10™wasemployedtotrackcellularapoptosisasafunctionofmitochondrialmembranepotential,andconsequently,mitochondrialactivity.Researcherswereinterestedinthepossibleanestheticpropertiesofmidazolamforanticancerdrugdelivery. - MitochondrialproteomicswithsiRNAknockdowntorevealACAT1andMDH2inthedevelopmentofdoxorubicin-resistantuterinecancer
JC-10™wasusedbyresearchersforthepurposesofdrugdiscovery.Inparticular,researcherswereinterestedinfindingnewtreatmentsfordoxorubicin-resistantuterinecancer,usingJC-10™tomonitormitochondrialmembranepotentialandvalidatecellviabilityresults. - Coldexposurelowersenergyexpenditureatthecellularlevel
ResearchersusedJC-10™toinvestigatetherelationshipbetweentemperatureandcellularactivity.Inparticular,researcherswantedtoexploreifcoldtemperatureactsasastressoronmitochondrialmembranepotential,withregardstotheoxidativephosphorylationprocesswhichgeneratesATP. - Susceptibilityofgametesandembryosoftheeasternoyster,Crassostreavirginica,toKareniabrevisanditstoxins
JC-10™wasusedinthestudyofspermviability,fertilizationsuccesssandembryonicsurvivalofCrassostreavirginica.Specifically,JC-10™wasusedtoquantifymitochondrialmembranepotentialinspermcellsandtodeterminepossibletoxicityeffectsofalgalblooms. - CalmodulinantagoNISTsinducecellcyclearrestandapoptosisinvitroandinhibittumorgrowthinvivoinhumanmultiplemyeloma
JC-10™wasusedbyresearcherstostudycellcycleandapoptosisinhumanmultiplemyeloma.JC-10™functionedasaprobeforthedetectionofmitochondrialmembranepotentialdepolarization,whichwascrucialtothestudyofcaspaseactivatedapoptosis. - ActivationofthemitochondrialapoptoticpathwayproducesreactiveoxygenspeciesandoxidativedamageinhepatocytesthatcontributetolivertumOrigenesis
Researcherswereinterestedinthepathwaysinvolvedwithlivertumorigenesis.Tothatend,theyusedJC-10™tostudyactivationofapoptoticpathwaysrelatedtochangesinmitochondrialmembranepotential,withtheultimategoalofdiscoveringifantioxidanttherapymighthelpsuppresslivercarinogenesis.