| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 510/525 |
| MW | ~600 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | CellAnalysis CellApoptosis |
| Related | VitalStains NeurologicalStains BiochemicalAssays |
- • ImprovedSolubility-lesslikelytoprecipitateinaqueoussolutions
- • GreaterConvenience-streamlinedprotocoltominimizehands-ontime
- • HigherQualitySignals-improvedsignaltobackgroundratio
- • MoreRobustAssay-optimizedprobeformoreconsistentresults
AssayPrinciple
JC-10™isacationic,lipophilicdyethatcanbeusedtomonitormitochondrialmembranepotential.Whenmitochondrialmembranepotentialislow,JC-10™emitsagreenfluorescence.Conversely,whenmitochondrialmembranepotentialishigh,JC-10™willemitagreen-orangefluorescence.
| Excitation | Emission | |
| LowMembranePotential | ~510nm | ~520nm |
| HighMembranePotential | ~510nm | ~570nm |
TheshiftinemissionisaresultofJC-10™"suniqueABIlitytoreversIBLyformaggregatesasafunctionofmitochondrialmembranepotential.Atlowmembranepotentials,JC-10™accumulationinmitochondriaislowandmostoftheJC-10™willbeinthemonomericform,whichwillemitat~520nmwhenexcitedby~490nmlightsource.Athighmembranepotentials,JC-10™accumulationinmitochondriaishighandmostoftheJC-10™willbeintheaggregateform,whichwillemitof~570nmwhenexcitedby~490nmlightsource.BecauseJC-10™aggregateformationisafunctionofmembranepolarization,thefluorescenceintensityofJC-10™aggregatesreflectsthedegreetowhichamitochondrialmembraneisdepolarizedorhyperpolarized.
Whilethereareolderprobesthatcanmonitormitochondrialmembranepotential,suchasJC-1,AATBioquest"sJC-10™outperformstheminseveralkeyaspects:
- UnlikeJC-1,whichtendstoprecipitateevenatlowconcentrations(ie.1µM),ourJC-10™probeismuchmoresolubleinaqueoussolutions.Thisallowsforeasierpreparationandhandling.
- OurJC-10™protocolhasbeenstreamlinedforminimalhands-ontime.Ourproductcomespreparedin2mg/mLsolution(ie.DMSO).Simplyadd,incubateandread.
- ExtensivetestinghasshowthatourJC-10™probehasbettersignalintensitythanotherprobes(seeFigure1)andhasahighersignaltobackgroundratio.
- Ourproductandprotocolshavebeenoptimizedtoensureconsistentresultsacrossmultipleruns.
- JC-10™hasbeendesignedforcompatibilitywithfluorescencemicroplatereaders,fluorescencemicroscopesandflowcytometersusingcommonFITC,TRITCandFL1/FL2channels(seeprotocolforadditionaldetails)
AdditionalFigures
| Spectrum | AdvancedSpectrumViewer |
1.PrepareJC-10workingsolution:
1.1 EachvialofDMSOstocksolution(100µL,2mg/mL,3mM)shouldbeusedonlyonce.Anyunusedvialsshouldbestoredat<-20oC.
Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
1.2 Preparea1XJC-10workingsolution:Onthedayoftheexperiment,thawanaliquotoftheJC-10stocksolutiontoroomtemperature.Preparea10to30µM1XworkingsolutioninHanksand20mMHepesbuffer(HHBS)orbufferofyourchoice,pH7-8with0.02%Pluronic®F-127.Mixthemwellbyvotexing.
Note:Forsomecelllines,workingsolutionatpH8mightpreventJC-10leakage.
2.RunJC-10assaywithafluorescencemicroplatereader:
2.1 Treatcellswithtestcompoundsforadesiredperiodoftime(Forexample,Jurkatcellscanbetreatedwithcamptothecinfor4-6hours)toinduceapoptosis.Forblankwells(mediumwithoutthecells),addthecorrespondingamountofcompoundbuffer.
2.2 Add100µL/well/96-wellplateor25µL/well/384-wellplateofJC-10workingsolution(fromStep1.2)intothecellplate.
2.3 IncubatetheJC-10loadingplateina37oC,5%CO2incubatorfor15-60min.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
2.4 MonitorthefluorescencechangeatEx/Em=490/525nm(FITCchannel)and540/595nm(TRITCchannel)forratioanalysis.
Optional:RemovetheJC-10workingsolutionfromtheplate;add100µL/well/96-wellplateor25µL/well/384-wellplateofHHBSbacktothecellplatebeforeanalysis.
3.RunJC-10assaywithafluorescencemicroscopeoraflowcytometer:
3.1 Treatcellswithtestcompoundsforadesiredperiodoftime(Forexample,Jurkatcellscanbetreatedwithcamptothecinfor4-6hours)toinduceapoptosis.
3.2 Centrifugethecellstoget1-5x105cellspertube.
3.3 ResUSPendcellsin500µLofJC-10workingsolution(fromStep1.2).
3.4 Incubateatroomtemperatureorina37°C,5%CO2incubatorfor10to30min,protectedfromlight.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
3.5 MonitorthefluorescencechangeatEx/Em=490/525nmand540/595nmwithafluorescencemicroscope(usingFITCandTRITCfilters)oraflowcytometer(usingFL1andFL2channels).
Optional:RemovetheJC-10workingsolutionfromtheplate;add100µL/well/96-wellplateor25µL/well/384-wellplateofHHBSbacktothecellplatebeforeanalysisonfluorescencemicroscope.
| 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.
AAT Bioquest AAT Bioquest是一家位于美国的生物公司，前身为ABD Bioquest，总部位于加利福尼亚州。专门从事光学检测技术十多年，一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术，综合了化学、生物学和信息学等各个领域的研究，引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络，为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。
美国AATBioquestInc.(前身是ABDBioquest，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。AATBioquest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。AATBioquest会不断介绍新产品，快速的丰富各个领域的产品。
1）我们提供反应荧光探针和发光探针，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）我们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）我们不断的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）我们致力于开发用于信号转导研究的试剂；5）我们提供生理和神经探针，特别是钙离子指示剂和膜电位探针。
作为AATBioquestInc.的中国区域代理，艾美捷科技为中国客户提供光谱学检测领域，包括吸收（颜色），荧光和发光技术等全系列解决方案。我们也将一如既往更加努力为国内用户提供快捷、方便的高质量产品，同时更为您售前售后全面技术支持。
AATBioquest,Inc.(formerlyABDBioquest,Inc.)develops,manufacturesandmarketsbioanalyticalresearchreagentsandkitstoscientistsengagedinlifesciencesresearch,diagnosticR&Danddrugdiscovery.Wespecializeintheareaofphotometricdetectionsincludingabsorption(color),fluorescenceandluminescencetechnologies.TheCompany"sproductsenablescientistsandbiomedicalresearcherstobetterunderstandbiochemistry,immunology,cellBIOLOGyandmolecularbiology.AATBioquestconstantlyintroducesnewproducts,andoffersarapidlyexpandinglistofproductsthataregroupedintoseveralproductlines.
1)Ourreactivefluorescentandluminescentprobes,biotinsandtagenzymesareusedforlabelingsmalldrugmoleculesandbiopolymers,e.g,proteins,nucleicacidsandcarbohydrates;2)Wedevelopfluorescentandluminescentprobesfordetectingproteins,nucleicacidsandlivecells;3)Weconstantlyintroducenovelfluorescentandluminescentprobesfordetectingvariousenzymes,inparticular,hydrolyticandredoxenzymes;4)Wefocusondevelopingreagentsforsignaltransductionresearch;and5)Wealsoofferphysiologicalandneurologicalprobes,e.g.,calciumindicatorsandmembranepotentialprobes.
Besidesthestandardcatalogproductswealsooffercustomservicetomeetyourspecialresearchneeds.Ourcurrentservicesincludecustomsynthesisofcolorimetric,fluorescentandluminescentprobes,customdevelopmentofbiochemical,cell-basedanddiagnosticassaysandcustomscreeningofyourcompoundlibrariesagainstyourdefinedtargetsusingourvalidatedHTSassays.
ebiomall.com
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请各位大侠给予帮助！！
谢谢！！
产品主要应用：点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
氨基类染料是包含自由氨基的活性染料，染料可与活化羧酸衍生物和其他亲电子的试剂结合。比如：氨基与EDC-活化的羧基结合。
相关产品如下：
中文名英文名产品编号分子结构Cy7.5胺Cy7.5amineAGF1350A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110804_5250.jpg[/img]Cy5胺Cy5amineAGF1332A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110715_7750.jpg[/img]相似系列产品：
抗体、核酸、蛋白质等生物分子标记染料
羰基活性荧光染料
巯基反应性染料
羧酸类染料
健那绿染液是一种活体染液，实验对象必须是活细胞，健那绿可以使活细胞中的线粒体呈现蓝绿色，而细胞质接近无色。
分子生物学实验的染料主要涉及到核酸染料和蛋白质染料.核酸染料主要有EB（溴化乙锭,高致癌性）,goldview,sybr green（实时定量PCR时常用染料）.这些染料可以和核酸双链分子特异性结合发出强荧光而被检测到.蛋白质染料最常用的是考马斯亮蓝 R-250,硝酸银.其中硝酸银有时也用于核酸染色.
染料分为天然染料和人工染料两种。天然染料有胭脂虫红、地衣素、石蕊和苏木素等，它们多从植物体中提取得到，其成分复杂，有些至今还未搞清楚。目前主要采用人工染料，也称煤焦油染料，多从煤焦油中提取获得，是苯的衍生物。多数染料为带色的有机酸或碱类，难溶于水，而易溶于有机溶剂中。为使它们易溶于水，通常制成盐类。
染料可按其电离后染料离子所带电荷的性质，分为酸性染料、碱性染料、中性（复合）染料和单纯染料四大类。 标本干燥后即进行固定，固定的目的有三个：
1）杀死微生物，固定细胞结构。
2）保证菌体能更牢的粘附在载玻片上，防止标本被水冲洗掉。
3）改变染料对细胞的通透性，因为死的原生质比活的原生质易于染色。
而且样品中的无水硫酸钠未变色,而做标准曲线的五个和空白对照的变为蓝色了,请高手指教,多谢!
还有,是否变蓝对测定结果有影响吗?
谢了哈
欢迎你！请下次规范发贴:)
产品主要应用：点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
菁染料是性能优良的荧光标记染料，摩尔吸光系数在荧光染料中是最高的。N-羟基琥珀酰亚胺酯是最常用的脂肪氨基标记试剂，广泛用于蛋白质、氨基酸多肽、抗体、核酸及其他生物分子的标记和检测。通过改变次甲基链的长度,可改变其荧光发射波长，每增加一个双键，按照Huoffman规则正好红移约100nm。
菁染料Cy3和Cy5已成为基因芯片的首选荧光标记物；另外,Cy5,Cy5.5和Cy7，Cy7.5的吸收在近红外区背景非常低，是荧光强度最高、最稳定的长波长染料，特别适合于活体小动物体内成像。但由于菁染料,尤其是不对称菁染料的合成副反应多,副产物极性相近,产物的分离提纯相当困难。菁染料特别是水溶性菁染料分子极性大,分离提纯越加困难。Lumiprobe供应脂溶性和水溶性菁染料。
相关产品：
产品分子结构可替代染料编号：AGF1371A
6-ROX-N-羟基琥珀酰亚胺酯
ROXNHSester,pure6-isomer[img]/KindEditor_4.0.1/attached/image/20130704/20130704102819_7906.jpg[/img]AlexaFluor568编号：AGF1326A
Cy3-N-羟基琥珀酰亚胺酯
Cy3NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103024_8531.jpg[/img]AlexaFluor546NHSester
DyLight549NHSester
编号：AGF1330A
Cy3.5-N-羟基琥珀酰亚胺酯
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DyLight594NHSester
编号：AGF1338A
Cy5-N-羟基琥珀酰亚胺酯
Cy5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103422_4468.jpg[/img]AlexaFluor647NHSester
DyLight649NHSester
编号：AGF1345A
Cy5.5-N-羟基琥珀酰亚胺酯
Cy5.5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103557_5875.jpg[/img]AlexaFluor680NHSester
DyLight680NHSester
编号：AGF1349A
Cy7-N-羟基琥珀酰亚胺酯
Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103713_8843.jpg[/img]编号：AGF1356A
Cy7.5-N-羟基琥珀酰亚胺酯
Cy7.5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103756_5406.jpg[/img]编号：AGF1374A
磺酸基-Cy3-N-羟基琥珀酰亚胺酯
Sulfo-Cy3NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103830_1656.jpg[/img]AlexaFluor546
DyLight549
编号：AGF1377A
磺酸基-Cy5-N-羟基琥珀酰亚胺酯
Sulfo-Cy5NHSester[img]/KindEditor_4.0.1/attached/image/20130705/20130705095752_6656.jpg[/img]AlexaFluor647
DyLight649
编号：AGF1379A
磺酸基-Cy7-N-羟基琥珀酰亚胺酯
Sulfo-Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704104047_6968.jpg[/img]相似系列产品：
羰基活性荧光染料
氨基类染料
巯基反应性染料
羧酸类染料
2、苏丹三 脂肪 橙红
3、苏丹四 脂肪 红
4、双缩脲 蛋白质 紫
5、龙胆紫 染色质 紫
6、碘 淀粉 蓝
7、健那绿 线粒体 绿
8、甲基绿 DNA 绿
9、吡罗红 RNA 红
10、溴麝香草酚蓝 CO2 由蓝变绿再变黄
11、重铬酸钾 酒精 酸性条件下由橙色变成灰绿
12、醋酸洋红（龙胆紫、改良苯酚品红） 染色质 红
13、台盼蓝 检验活死细胞 死细胞会被染成蓝色（不常用）
