ReadiUse™ Bio-Gel P-6 spin column | AAT Bioquest

Invert the Spin Column several times to suspend the settled gel and remove any bubbles. Snap off the tip and place the column in a microcentrifuge tube (2 mL). Activelyhelpingcustomers,employeesandtheglobalcommunityduringthecoronavirusSARS-CoV-2outbreak. Learnmore>>XNavigationMenuHomeSignInShoppingCartOrderingInformationProductsServicesToolsResourcesAboutUsCareersContactUsDistributorsTermsofUseTermsofSalesPrivacyAATBioquest ReadiUse™Bio-GelP-6spincolumnOrderinginformationPrice()CatalogNumber60500UnitSize5FindDistributorAdditionalorderinginformationTelephone1-408-733-1055Fax1-408-733-1304Emailsales@aatbio.comInternationalSeedistributorsShippingStandardovernightforUnitedStates,inquireforinternationalStorage,safetyandhandlingIntendeduseResearchUseOnly(RUO)StorageRoomtemperature(10-25°C)OverviewSDSProtocolSeealso:BuffersandLabConsumablesCAS25034-58-6ReadiUse™SpinchromatographycolumnsarepackedwithpolyacrylamideP-6-DGmatricesforbufferexchangeanddesaltingapplications.Whenapplyingamaterialtothespincolumn,thesubstancessmallerthanthecolumn'sexclusionlimit(MW~6000)willberetainedinthecolumnandlargemoleculeslikeproteins(MWisabove15,000)willbeelutedwithabuffer.Thesamplevolumerangesfrom30uLto100uL.Theyarecompatiblewithswingingbucketcentrifugewith2.0mlmicrocentrifugetubesor12x75testtubes.ExampleprotocolSAMPLEEXPERIMENTALPROTOCOL InverttheSpinColumnseveraltimestosuspendthesettledgelandremoveanybubbles. Snapoffthetipandplacethecolumninamicrocentrifugetube(2mL).Removethecaptoallowtheexcesspackingbuffertodrainbygravityuntilreachingthetopofthegelbed.Ifcolumndoesnotbegintoflow,pushcapbackintocolumnandremoveitagaintostarttheflow.Discardthedrainedbuffer,andthenplacethecolumnbackintotheTube. Centrifugefor1mininaswingingbucketcentrifugeat1,000xgtoremovethepackingbuffer.Discardthebuffer. Apply1mLequilibriumbufferofyourchoicetothecolumn,letthebufferdrainoutbygravity,orcentrifugethecolumnfor1mintoremovethebuffer.Discardthebufferfromthecollectiontube.Repeatthisprocessfor3-4times. Centrifugefor2minutesinaswingingbucketcentrifugeat1,000xgtoremovethereactionbuffer.Discardthebuffer. Placethecolumninaclean2.0mlmicrocentrifugetubeor12x75mmtesttube.Carefullyapplythesample(20–100µl)directlytothetopcenterofthecolumn.Samplevolumemayneedbecarefullyadjustedtoreachthebestperformance. Afterloadingsample,centrifugethecolumnfor5minat1,000xg. Followingcentrifugation,thepurifiedsampleisnowintheequilibriumbufferandmoleculessmallerthanthecolumn’sexclusionlimit(~6000)willberetained. Collecttheelution,andproperlydisposetheusedcolumn.Note:SpinColumncanfitinto2mLmicrocentrifugetubesor12x75mmtesttubesforsamplecollectionduringcentrifugation.Usethe2mLmicrotubesprovidedwiththecolumnsfortheinitialcolumnequilibrationstep.Note:Swingingbucketcentrifugescapableofgeneratingaminimumforceof1,000xgaresuitableforBio-Spincolumnuse.Thegravitationalforcecreatedataparticularrevolutionspeedisafunctionoftheradiusofthemicrocentrifugerotor.Consulttheswingingbucketcentrifugeinstructionmanualfortheinformationaboutconversionfromrevolutionsperminute(RPM)tocentrifugalorg-force.Alternatively,usetheequationshownbelowtocalculatethespeedinRPMrequiredreachingthegravitationalforceof1,000xg.                              CF(g)=(1.12X10-5)X(RPM)2Xr          RCF=therelativecentrifugalforce,RPM=thespeedoftherotorr=theradiusincentimetersmeasuredfromthecenteroftherotortothemiddleoftheBio-SpincolumnImagesFigure1.PrepackedReadiUse™Bio-GelP6SpinColumn.ThecolumnwaspackedwithP-6DGin PBSbufferforsamplevolume50~100uL.TheMWlimitis~6000andlargemoleculeslikeproteins(MW >15,000)willbeelutedwithabuffer.DocumentsSafetydatasheetProductprotocolCertificateofanalysisFAQHowdoyoupermeabilizeacell?WhatarethestepsofDNAextraction?Whatisthebestwaytomakeyourownlysisbufferforproteinisolation?Whichcelllysisbufferrecipeisbestforphosphorylatedproteins?AssayWisePortelite™FluorimetricQuantitationKits:CompanionReagentsOptimizedfortheCytoCite™BG100SpectrofluorometerLocation:Home/Products/ReadiUse™Bio-GelP-6spincolumnAboutPrivacyTermsofUseTermsofSalesDistributorsCopyright©2021AATBioquest,Inc.AllRightsReserved.