PETCellDissociationReagentisacelldissociationmixspeciallyformulatedforusewiththe"ColdTrypsin"method.Whenusedtodetachmonolayerculturesofepithelialcellsfromthegrowthsubstratum,thisreagentwillyieldmoreviablecellsofmonolayersgrowninserum-freemedium.Thesolutioncontainspolyvinylpyrrolidone,EGTAandtrypsininaHBSbase.||SuggestedUse:|AfterrinsingthecellmonolayersusingHBS,add3mloficecoldPETdissociationreagent.Donotkeeptheflaskintheincubator.Incubateatroomtemperatureandmonitorthemonolayerbymicroscopy(preferablyphasecontrast).Whenthecellsbegintoseparate,gentlyremovemostofthePET(withoutremovingthecells)andallowtheflasktoremainatroomtemperature.Todeterminewhetherthecellsarereadytoharvest,taptheflask(heldinanuprightposition)firmlybyhand.||Whenthecellsfloatfree,resUSPendthemincoldHBSsupplementedwith5%FetalBovineSerumtostopthetrypticaction.Mixwellbypipettingthecellsuspension,centrifugeandresuspendthecellpelletintheappropriatecompleteserum-freemedium.||pH:7.2-7.6(5%CO2atmosphereisrecommendedforculture.)||Conductivity:|Asreported||TrypsinActivity:DeterminedusingPDQTMProteaseAssay.||SterilityTest:|Nodetectionofmicrobialgrowth.||StorageandStABIlity:|Maybestoredat4°Cforshort-term(4-6weeks).Forlong-termstorage,aliquottoavoidrepeatedfreezingandthawingandfreezeat-70°C.Aliquotsarestablefor6months.

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