Overview:
Abetterwaytoisolateexosomes
“WethereforepursuedtheExoQuick®methodforfurtherstudy,asthesesamplesrequiredmuchlesssampleinput,akeybenefitwhenworkingwithclinicalsamplesandmousemodels1.”
Needexosomes?SBI’sExoQuick-TCenableshigh-throughput,quantitativeisolationofexosomesfromlowvolumes(aslittleas1ml)ofmostbiofluids(forserum,plasma,orascitesfluid,usetheoriginalExoQuickformulation(EXOQ5A-1orEXOQ20A-1)).CompatIBLewithawidevarietyofdownstreamapplications,ExoQuick-TCisaneffectiveandprovenalternativetoultracentrifugation1-3.
ExoQuick-TC’sfast,ultracentrifugation-freemethod:
- Savestimeandlabor
- Iseasilyscalable
- Conservesprecioussample
- Delivershighyieldsoffunctional,highqualityexosomes
- Canbeusedtoisolateexosomesforawiderangeofdownstreamapplications,including
ExoQuick-TCisaproprietarypolymerthatgentlyprecipitatesexosomes.First,pre-clearyoursamplesofcellsandcellulardebris,andthensimplyaddtheappropriateamountofExoQuick-TCtoyourclearedbiofluid,refrigerate,andcentrifuge(seetheproductmanualforprotocoldetails).Yourexosomeswillbeinthepellet,readyforresUSPensioninanappropriatesolution.
| Biofluid | Samplevolume | ExoQuick-TCVolume |
|---|---|---|
| Tissueculturemedia,urine,cerebrospinalfluid(CSF),etc. | 5mL | 1mL |
Inelectronmicroscopystudies,exosomesisolatedwithExoQuick-TCappearsimilartoexosomesisolatedusingultracentrifugation1-2,andtheseexosomesarealsoactiveinnumerousfunctionalassays1-3.
ExosomesisolatedwithExoQuick-TCcanbeusedforalltypesofproteinprofilingandproteincharacterizationstudies,suchasmassspectrometry,Westernblotting,ELISA,andmore.HigherproteinyieldsareachievedbyExoQuickpurificationthanbychromatography,DynaBeads,orultracentrifugation.
ExosomesisolatedwithExoQuick-TCalsoprovideexcellentsamplesforstudyingexosome-associatednucleicacidssuchasmicroRNAs,siRNAs,andevenmRNA.QuantitativeanalyticaltechniquessuchasqPCR,microarraystudies,andnext-generationsequencingareallcompatiblewithnucleicacidsisolatedfromExoQuick-TC-purifiedexosomes.
Backedbyagrowingnumberofpublications,ExoQuick-TCisoftenthebestoptionforresearchersworkingwithlowsamplevolumes,suchasclinicalresearchsamplesorsmallanimalmodels.
ExoQuick-TCexosomeisolationmethodsarepatentedtechnologies4.
ChoosetherightExoQuickforyourbiofluid:
| Catalog# | Product | Biofluid |
|---|---|---|
| EXOQ5A-1,EXOQ20A-1 | TheOriginalExoQuick | Forserum,plasma,andascitesfluid |
| EXOTC10A-1,EXOTC50A-1 | ExoQuick-TC® | Forallotherbiofluids |
| EXOLP5A-1 | ExoQuick-LP | ForremovingcontaminatinglipoproteinparticlesfromplasmaorserumbeforeExoQuickprecipitation |
| EXOQ5TM-1 | ExoQuickPlasmaPrepwithThrombin | Forde-fibrinatingplasmabeforeexosomeisolationforefficientrecoveryandhighyields |
| EXOCG50A-1 | ExoQuick-CG | Forpreparingexosomesusedinpre-clinicalinvivoapplications |
| EQPL10A-1,EQPL10TC | ExoQuickPLUSandExoQuick-TCPLUS | Forsensitiveapplicationssuchasmassspectrometry,exosomelabeling,andinvivo/exvivoexosomedelivery |
ChoosebetweenExoQuick/ExoQuick-TCandExoQuickPLUS/ExoQuick-TCPLUSbasedonyourapplication
| ExoQuick ExoQuick-TC | ExoQuickPLUS ExoQuick-TCPLUS | |
ProteinDetection | ||
| Westernblottingforgeneralexosomemarkers(e.g.CD9,CD63,CD81,TSG101,Alix) | ••••• | ••••• |
| High-sensitivityWesternblotting(e.g.lowabundancebiomarkers) | ••• | ••••• |
qPCRAnalysis | ||
| qPCRofcodingandnon-codingRNAs(e.g.mRNA,miRNA,andlncRNA) | ••••• | ••••• |
High-throughputBiomarkerDiscovery | ||
| RNA-seqofexosomalRNAs | ••••• | ••••• |
| Massspectrometryofexosomalproteins | ••• | ••••• |
| Lipidomics/metabolomicsofexosomalcargo | ••• | ••••• |
ExosomeLabeling | ••• | ••••• |
| Invivo/exvivoExosomeDelivery | ••• | ••••• |
•••••Highlyrecommended,•Notrecommended | ||
REFERENCES
- ChughPE,etal.SystemicallyCirculatingViralandTumor-DerivedMicroRNAsinKSHV-AssociatedMalignancies.PLoSPathog.2013.9(7):e1003484.PMCID:PMC3715412.
- UmezuT,etal.LeukemiacelltoendothelialcellcommunicationviaexosomalmiRNAs.Oncogene.2013May30.32(22):2747-55.PMID:22797057.
- SohelMM,etal.ExosomalandNon-ExosomalTransportofExtra-CellularmicroRNAsinFollicularFluid:ImplicationsforBovineOocyteDevelopmentalCompetence.PLoSOne.2013Nov4.8(11):e78505.PMCID:PMC3817212.
- AntesT,etal.MethodsforMicrovesicleIsolationandSelectiveRemoval.PatentNo.:US9,005,888B2.
HowItWorks:
High-throughput,quantitativeexosomerecovery
ExoQuick-TCcanbeusedtopurifyexosomesfromawidevarietyofbiofluids,includingsaliva1,urine2,follicularfluid3,tissueculturemedia4,andbreastmilk5.Withasimpleworkflowinvolvingminimalhands-ontimeandlowinputsamplevolumerequirements,ExoQuick-TCisanexcellentoptionforresearcherswhoneedtopurifymultipleexosomesamples.
Toisolateexosomesfromanyclearedbiofluid,simply:
AddanappropriatevolumeofExoQuick-TCIncubateforatleastonehourat4°CIsolateexosomeswitha30-minutelow-speedspin(1500g).Isolatedexosomescanbefoundinthepelletandresuspendedinanappropriatesolution.
Youcanverifythepresenceofexosomeswithanumberofdifferentmethods,includingWesternblottingforgeneralexosomemarkers(CD63,CD9,CD81,andHSP70),NanoSightanalysis,orEM(learnaboutdifferentwaystodetectexosomesandmoreinourExosomeBasicsGuide).
TheBottomLine
WithExoQuick-TC,youcanobtainhigh-qualityexosomesfrommostbiofluidsusingaprotocolthatcaneasilybeperformedonmultiplesamplesandrequiresverylowvolumesofinputsample.
REFERENCES
YangJ,etal.DetectionofTumorCell-SpecificmRNAandProteininExosome-LikeMicrovesiclesfromBloodandSaliva.PLoSONE.2014.9(11):e110641.PMCID:PMC4232306.AlvarezML.IsolationofurinaryexosomesforRNAbiomarkerdiscoveryusingasimple,fast,andhighlyscalablemethod.MethodsMolBiol.2014.1182:145-70.PMID:25055908.SohelMM,etal.ExosomalandNon-ExosomalTransportofExtra-CellularmicroRNAsinFollicularFluid:ImplicationsforBovineOocyteDevelopmentalCompetence.PLoSOne.2013Nov4.8(11):e78505.PMCID:PMC3817212.ZhuL,etal.Novelmethodforextractingexosomesofhepatocellularcarcinomacells.WorldJGastroenterol.2014June7.20(21):6651-6657.PMCID:PMC4047354.GuY,etal.Lactation-RelatedMicroRNAExpressionProfilesofPorcineBreastMilkExosomes.PLoSONE.2012.7(8):e43691.PMCID:PMC3427246.Citations:
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- Maeda,Y,etal.(2017)Synovium-DerivedMicroRNAsRegulateBonePathwaysinRheumatoidArthritis.J.BoneMiner.Res..2017Mar1;32(3):461-472.PMID:27676131
- Umezu,T,etal.(2017)ReplenishingexosomesfromolderbonemarrowstromalcellswithmiR-340inhibitsmyeloma-relatedangiogenesis.BloodAdvances.2017May16;1(13):812-823.Link:BloodAdvances
- Saha,B,etal.(2017)CirculatingandExosome-PackagedHepatitisCSingle-StrandedRNAInduceMonocyteDifferentiationviaTLR7/8toPolarizedMacrophagesandFibrocytes.J.Immunol..2017Mar1;198(5):1974-1984.PMID:28122964
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- Huang,Z,etal.(2017)UrinaryExosomalmiR-193aCanBeaPotentialBiomarkerfortheDiagnosisofPrimaryFocalSegmentalGlomerulosclerosisinChildren.BiomedResInt.2017Mar1;2017:7298160.PMID:28246603
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