G25脱盐柱使用指南

DesaltingcolumnprotocolMr.YuWang1精选2021版课件MatrixSephadex™G-25Fine,cross-linkeddextranBedvolume53mlBedheight100mmi.d.26mmseparationofhigh(Mr>5000)fromlowmolecularweightsubstances(Mr<1000).Storage+4to+30°20%ethanol2精选2021版课件长期不用后再使用，应该用5CV平衡柱子（5*60=300mlbuffer）TrytheseconditionsfirstBuffer(35ms/cm):50mMphosphatebuffer,0.15MNaCl,pH7.0Flowrate:20ml/minatroomtemperatureSamplevolume:15mlSeparations:Elutethecolumnwith106ml(2CV)ofbuffer3精选2021版课件Equilibrationisnotnecessarybetweenrunswiththesamebuffer.(去除气泡）Reversetheflowdirectionandpump200mlwellde-gassedbufferataflowrateof15ml/minthroughthecolumn.Thequalityofthepackedbedwillnotnormallybeaffected.4精选2021版课件5精选2021版课件6精选2021版课件7精选2021版课件SephadexG-10iswellsuitedfortheseparationofbiomoleculessuchaspeptides(Mr>700)fromsmallermolecules(Mr<100).SephadexG-50issuitablefortheseparationofmoleculesMr>30000frommoleculesMr<1500suchaslabeledproteinorDNAfromunconjugateddyes.Themediumisoftenusedtoremovesmallnucleotidesfromlongerchainnucleicacids.SephadexG-25isrecommendedforthemajorityofgroupseparationsinvolvingglobularproteins.ThesemediaareexcellentforremovingsaltandothersmallcontaminantsawayfrommoleculesthataregreaterthanMr5000.8精选2021版课件samplevolumesofupto30%ofthetotalvolumeofthedesaltingcolumncanbeprocessed.notexceedapproximately70mg/ml。9精选2021版课件Column:HiTrapDesalting,1×5ml,3×5ml,5×5mlSample:2mg/mlBSAin50mMsodiumphosphate,0.5Msodiumchloride,pH7.0Samplevolume:28%×Vt(1.4,4.3and7.1mlrespectively)Buffer:50mMsodiumphosphate,0.15Msodiumchloride,pH7.0Flowrate:5ml/min10精选2021版课件StorageStoreunusedmedia4°Cto30°Cin20%ethanol.Donotfreeze.Washusedmediawith2columnvolumesofdistilledwaterfollowedby2columnvolumesof20%ethanol.Storeat4°Cto30°C.Alternatively,washwith2columnvolumesofdistilledwaterfollowedby2columnvolumes0.01MNaOH.Sodiumhydroxidesolutionisbacteriostatic,easilydisposedofanddoesnotshrinkthemedium.Degastheethanol/watermixturethoroughlyandusealowflowrate,checkingtheback