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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*/22801/
商品详细AAT Bioquest/Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*/22801/
AAT Bioquest/Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*/22801/
AAT Bioquest/Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*/22801/
商品编号: 22801
品牌: aatbio
市场价: ¥85560.00
美元价: 51336.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)510/525
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
BiochemicalAssays
AlthoughJC-1iswidelyusedinmanylabs,itspoorwatersolubilitycausesextraordinaryinconvenience.Evenat1µMconcentration,JC-1tendstoprecipitateinaqueousbuffer.JC-10isdevelopedtobeasuperioralternativetoJC-1wherehighdyeconcentrationisdesired.ComparedtoJC-1,JC-10hasmuchbetterwatersolubility.JC-10iscapableofenteringselectivelyintomitochondria,andchangesreversIBLyitscolorfromgreentoorangeasmembranepotentialsincrease.ThispropertyisduetothereversibleformationofJC-10aggregatesuponmembranepolarizationthatcausesshiftsinemittedlightfrom520nm(i.e.,emissionofJC-10monomericform)to570nm(i.e.,emissionofJ-aggregate).Whenexcitedat490nm,thecolorofJC-10changesreversiblyfromgreentogreenishorangeasthemitochondrialmembranebecomesmorepolarized.Bothcolorscanbedetectedusingthefilterscommonlymountedinallflowcytometers,sothatgreenemissioncanbeanalyzedinfluorescencechannel1(FL1)andgreenishorangeemissioninchannel2(FL2).Besidesitspotentialuseinflowcytometry,itcanalsobeusedinfluorescenceimagingandfluorescencemicroplateplatform.Thiskitprovidesalltheessentialcomponentswithanoptimizedassaymethodforthedetectionofapoptosisincellswiththelossofmitochondrialmembranepotential.Thisfluorometricassayisbasedonthedetectionofthemitochondrialmembranepotentialchangesincellsbythecationic,lipophilicJC-10dye.Innormalcells,JC-10concentratesinthemitochondrialmatrixwhereitformsredfluorescentaggregates.However,inapoptoticandnecroticcells,JC-10existsinmonomericformandstainscellsingreenfluorescence.Thekitisoptimizedforscreeningofapoptosisactivatorsandinhibitorsbyflowcytometry.Wealsoofferaconvenient96-welland384-wellfluorescencemicrotiter-plateformatkit(cat#22800)forhighthroughputscreening.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

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
JC-10™hasbeenusedtostudymanyBIOLOGicallysignificantprocessesacrossseveralkeydisciplines.Tolistafew,JC-10™hasbeenusedtoinvestigatetopicssuchasmitochondrialmembranepotential,cytotoxicity,cellviABIlity,oxidativestress,cancermetastasis,apoptosis,signaltransduction,mitochondrialfissionandinducedpluripotentstemcells(iPSCs).

Below,youmayfindasmallsamplingofspecificJC-10™applications.ToinquireaboutapotentialapplicationofJC-10™,ortoconsultwithourfluorescentdyespecialists,pleasecontactusatsupport@AATbio.comor1-800-990-8053.

  1. High-contentassaysforhepatotoxicityusinginducedpluripotentstemcell–derivedcells.
    ResearchersuseJC-10™tomonitormitochondrialdepolarizationasanindicationofhepatotoxicityandoxidativestressininducedpluripotentstemcell-derivedcells,withtheultimategoalofdesigningareliable,high-contentandimaging-basedinvitrotoxicityassay.

  2. High-ContentHigh-ThroughputAssaysforCharacterizingtheViabilityandMorphologyofHumaniPSC-DerivedNeuronalCultures
    JC-10™wasusedtostudyneuronsderivedfrominducedpluripotentstem-cells.SinceJC-10™willaccumulateinthemitochondriaofviablecells,itwasusedtodeterminecellviabilityinahigh-throughputassaycontext.

  3. AnticancerActivityofNewSyntheticα-Methylene-δ-LactonesonTwoBreastCancerCellLines
    ResearcherschoseJC-10™toinvestigatemitochondrialmembranepotentialandmembraneintegrityincellstreatedwithnaturalproducts,suchasα-Methylene-δ-Lactones,withthegoalofdevelopingnewtreatmentsforbreastcancer.

  4. Tetrandrineprotectsmouseretinalganglioncellsfromischemicinjury.
    JC-10™wasusedinflowcytometrytostudymitochondrialmembranepotential(ΔΨm)inprimaryculturedretinalganglioncells,asanextensionofthefieldofdrugdiscoveryintopreventionofischemicinjury.

  5. Midazolaminducescellularapoptosisinhumancancercellsandinhibitstumorgrowthinxenograftmice
    Inastudyofhumancancercells,JC-10™wasemployedtotrackcellularapoptosisasafunctionofmitochondrialmembranepotential,andconsequently,mitochondrialactivity.Researcherswereinterestedinthepossibleanestheticpropertiesofmidazolamforanticancerdrugdelivery.

  6. MitochondrialproteomicswithsiRNAknockdowntorevealACAT1andMDH2inthedevelopmentofdoxorubicin-resistantuterinecancer
    JC-10™wasusedbyresearchersforthepurposesofdrugdiscovery.Inparticular,researcherswereinterestedinfindingnewtreatmentsfordoxorubicin-resistantuterinecancer,usingJC-10™tomonitormitochondrialmembranepotentialandvalidatecellviabilityresults.

  7. Coldexposurelowersenergyexpenditureatthecellularlevel
    ResearchersusedJC-10™toinvestigatetherelationshipbetweentemperatureandcellularactivity.Inparticular,researcherswantedtoexploreifcoldtemperatureactsasastressoronmitochondrialmembranepotential,withregardstotheoxidativephosphorylationprocesswhichgeneratesATP.

  8. Susceptibilityofgametesandembryosoftheeasternoyster,Crassostreavirginica,toKareniabrevisanditstoxins
    JC-10™wasusedinthestudyofspermviability,fertilizationsuccesssandembryonicsurvivalofCrassostreavirginica.Specifically,JC-10™wasusedtoquantifymitochondrialmembranepotentialinspermcellsandtodeterminepossibletoxicityeffectsofalgalblooms.

  9. CalmodulinantagoNISTsinducecellcyclearrestandapoptosisinvitroandinhibittumorgrowthinvivoinhumanmultiplemyeloma
    JC-10™wasusedbyresearcherstostudycellcycleandapoptosisinhumanmultiplemyeloma.JC-10™functionedasaprobeforthedetectionofmitochondrialmembranepotentialdepolarization,whichwascrucialtothestudyofcaspaseactivatedapoptosis.

  10. ActivationofthemitochondrialapoptoticpathwayproducesreactiveoxygenspeciesandoxidativedamageinhepatocytesthatcontributetolivertumOrigenesis
    Researcherswereinterestedinthepathwaysinvolvedwithlivertumorigenesis.Tothatend,theyusedJC-10™tostudyactivationofapoptoticpathwaysrelatedtochangesinmitochondrialmembranepotential,withtheultimategoalofdiscoveringifantioxidanttherapymighthelpsuppresslivercarinogenesis.


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品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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