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1Amicon®Ultra-1510KCentrifugalFilterDevicesEnglishIntroductionAmicon®Ultra-1510Kcentrifugalfilterdevicesprovidefastultrafiltration,withthecapabilityforhighconcentrationfactorsandeasyconcentraterecoveryfromdiluteandcomplexsamplematrices.Theverticaldesignandavailablemembranesurfaceareaprovidefastsampleprocessing,highsamplerecovery(typicallygreaterthan90%ofdilutestartingsolution),andthecapabilityfor80-foldconcentration.Typicalprocessingtimeis15to40minutes.Solutepolarizationandsubsequentfoulingofthemembraneareminimizedbytheverticaldesign,andaphysicaldeadstopinthefilterdevicepreventsspinningtodrynessandpotentialsampleloss.Theconcentrateiscollectedfromthefilterdevicesamplereservoirusingapipettor,whiletheultrafiltrateiscollectedintheprovidedcentrifugetube.Thedevicecanbespuninaswinging-bucketorfixed-anglerotor.Amicon®Ultra-1510Kdevicesaresuppliednon-sterileandareforsingleuseonly.IntendedUseTheAmicon®Ultra-15productlineincludes5differentcutoffs(MolecularWeightCutoff,MWCO),however,theAmicon®Ultra-1510Kdevice(10,000MWCO)istheonlydeviceintendedforinvitrodiagnosticuse.Itcanbeusedtoconcentrateserum,urine,cerebrospinalfluid,andotherbodyfluidspriortoanalysis.ForinformationonotherAmicon®Ultra-15cutoffs,goto Ultra-15inthesearchbox.UserGuideAmicon®Ultra-1510KCentrifugalFilterDevicesforvolumesupto15mLUFC901008UFC901024UFC901096CV2Amicon®Ultra-1510KCentrifugalFilterDevicesApplications●Concentrationofbiologicalsamplescontainingantigens,antibodies,enzymes,nucleicacids(DNA/RNAsamples,eithersingle-ordouble-stranded),microorganisms,columneluates,andpurifiedsamples●Purificationofmacromolecularcomponentsfoundintissuecultureextractsandcelllysates,removalofprimer,linkers,ormolecularlabelsfromareactionmix,andproteinremovalpriortoHPLC●Desalting,bufferexchange,ordiafiltrationMaterialsSuppliedTheAmicon®Ultra-1510Kdeviceissuppliedwithacap,afilterdevice,andacentrifugetube.FilterdeviceCentrifugetubeCapRequiredEquipment●Centrifugewithswinging-bucketorfixed-anglerotorwithwells/carriersthatcanaccommodate50 mLtubesCAUTION:Toavoiddamagetothedeviceduringcentrifugation,checkclearancebeforespinning.●Pipettorwith200microliter(μL)tipforconcentraterecoverySuitabilityPreliminaryrecoveryandretentionstudiesaresuggestedtoensuresuitabilityforintendeduse.Seethe“HowtoQuantifyRecoveries”section.ShelfLifeShelflifeis3yearsfromdateofmanufacture.Refertoexpirationdateonpackagelabel.3Amicon®Ultra-1510KCentrifugalFilterDevicesRinsingBeforeUseTheultrafiltrationmembranesinAmicon®Ultra-1510Kdevicescontaintraceamountsofglycerine.Ifthismaterialinterfereswithanalysis,rinsethedevicewithbufferorMilli-Q®waterbeforeuse.Ifinterferencecontinues,rinsewith0.1NNaOHfollowedbyasecondspinofbufferorMilli-Q®water.CAUTION:DonotallowthemembraneinAmicon®Ultrafilterdevicestodryoutoncewet.Ifyouarenotusingthedeviceimmediatelyafterrinsing,leavefluidonthemembraneuntilthedeviceisused.HowtoUseAmicon®Ultra-15CentrifugalFilterDevices1.Addupto15mLofsample(12mLifusingafixed-anglerotor)totheAmicon®Ultrafilterdevice.2.Placecappedfilterdeviceintocentrifugerotor;counterbalancewithasimilardevice.3.Whenusingaswinging-bucketrotor,spinthedeviceat4,000×gmaximumforapproximately15–40 minutes.Whenusingafixed-anglerotor,orientthedevicewiththemembranepanelfacingupandspinat5,000×gmaximumforapproximately15–40minutes.NOTE:RefertoFigure1andTable1fortypicalspintimes.4.Torecovertheconcentratedsolute,insertapipettorintothebottomofthefilterdeviceandwithdrawthesampleusingaside-to-sidesweepingmotiontoensuretotalrecovery.Theultrafiltratecanbestoredinthecentrifugetube.NOTE:Foroptimalrecovery,removeconcentratedsampleimmediatelyaftercentrifugation.DesaltingorDiafiltrationDesalting,bufferexchange,ordiafiltrationareimportantmethodsforremovingsaltsorsolventsinsolutionscontainingbiomolecules.TheremovalofsaltsortheexchangeofbufferscanbeaccomplishedintheAmicon®Ultra-15devicebyconcentratingthesample,thenreconstitutingtheconcentratetotheoriginalsamplevolumewithanydesiredsolvent.Theprocessof“washingout”canberepeateduntiltheconcentrationofthecontaminatingmicrosolutehasbeensufficientlyreduced.Seeexamplebelow.SpintoconcentrateSpintoconcentrate15mLof1mg/mLproteinin100mMNaCl200μLof75mg/mLproteinin100mMNaCl100mMNaCl200μLof75mg/mLproteinin11.2mMNaCl11.2mMNaClAdd14.8mLof10mMNaClorexchangebuffer15mLof1mg/mLproteinin11.2mMNaCl4Amicon®Ultra-1510KCentrifugalFilterDevicesPerformanceFlowRateFactorsaffectingflowrateincludesampleconcentration,startingvolume,chemicalnatureofsolute,relativecentrifugalforce,centrifugerotorangle,membranetype,andtemperature.Figure1andTable1canbeusedtoestimatethetimerequiredtoachieveagivenvolumeoffiltrateorconcentrate.Atypicalspintimefora15mLsampleisapproximately15to40 minutes.Whilemostofthesampleisfilteredinthefirst15to30minutesofcentrifugation,thelowestconcentratevolume(100–150μL)isreachedafterspinningfor15to40minutes
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