1、GE Healthcare?Gel FiltrationPrinciples and Methods 18-1022-18Recombinant Protein Purification HandbookPrinciples and Methods 18-1142-75Protein PurificationHandbook18-1132-29Hydrophobic Interaction and Reversed Phase ChromatographyPrinciples and Methods 11-0012-692-D Electrophoresisusing immobilized
2、pH gradientsPrinciples and Methods 80-6429-60Microcarrier Cell CulturePrinciples and Methods 18-1140-62GST Gene Fusion SystemHandbook18-1157-58Affinity ChromatographyPrinciples and Methods18-1022-29Antibody PurificationHandbook18-1037-46PercollMethodology and Applications 18-1115-69Ion Exchange Chro
3、matography&ChromatofocusingPrinciples and Methods11-0004-21PurifyingChallenging ProteinsPrinciples and Methods28-9095-31Handbooksfrom GE HealthcareChallenging?4?.7?.11?.11?.14?.21?.21?.21?KTADESIGN?.22?.25?.25?.25?.25?NI SEPHAROSE HIGH PERFORMANCE?.34?NI SEPHAROSE 6 FAST FLOW?.38?HIS MULTITRAP HP?HI
4、S MULTITRAP FF 96?.44?HIS SPINTRAP?.48?HISTRAP HP?HISTRAP FF?.51?HISTRAPFF?KTAPRIME PLUS?.58?HISTRAP FF CRUDE?.62?HISTRAP FF CRUDE?.68?HIS GRAVITRAP?HIS GRAVITRAP KIT?.73?HISPREP FF16/10?.77?.79?IMAC SEPHARSOE HIGH PERFORMANCE?.81?IMAC SEPHARSOE 6 FAST FLOW?.84?HITRAP IMAC HP?HITRAP IMAC FF?.87?HIPR
5、EP IMAC FF 16/10?.91?.94?5?.97?.99?.103?.103?.103?.106?.109?GST?.111?.111?.111?.114?GST?.115?GST?.118?.119?GLUTATHIONE SEPHAROSE HIGH PERFORMANCE?GLUTATHIONE SEPHAROSE 4 FAST FLOW?GLUTATHIONE SEPHAROSE 4B?.120?GST MULTITRAP FF?GST MULTITRAP 4B 96?.127?GST SPINTRAP?.132?GSTRAP HP?GSTRAP FF?GSTRAP 4B?
6、.133?GSTPREP FF 16/10?.141?.145GST?.148?.151?GST?.153?.174?MBP?.176?MBPTrap HP?.176?.186?Strep-tag II?.188?StrepTactin Sepharose High Performance?.1886?StrepTrap HP1?5?.191?.198?.200?.205?.214?.214?.215?.219?.220PD-10 DESALTING.222HITRAP DESALTING.223HIPREP 26/10 DESALTING.223?1 NI SEPHAROSE?IMAC SE
7、PPHAROSE?.229NI SEPHAROSE?.229?IMAC SEPHAROSE?.237?2 GLUTATHIONE SEPHAROSE?.242?3?.247?4?.251?5?.254?6?CM/H?ML/MIN?.255?7 GST?.256PGEX?.257?8?.258?9?.2597?SpinTrapTM?MultiTrapTM?96?1?GST?2?1?Gel filtrationHydrophobic interactionIon exchangeAffinityReversed phase8?A280?280?AC?BCA?bicinchoninic acidCD
8、NB?1-chloro-2,4-dinitrobenzeneCF?CV?DAB?3,3-diaminobenzidineDNase?ELISA?FF?Gua-HCl?GF?GST?HIC?HMW?HP?HRP?IEX?IMAC?IPTG?-?-D-?LMW?MPa?Mr?N/m?PBS?pI?pH?psi?PMSF?PVDF:?r?rGST?rBCARNase?RPC?SDS:?SDS-PAGE?TCEP?(2-?)?9?10?2?NativeconditionsBinding buffer(including 20 to40 mM imidazole)Binding buffer(inclu
9、ding 20 to40 mM imidazole)DenaturingconditionsCell lysisBinding toaffinity mediaWashElutePure denaturedtagged proteinPure taggedproteinPure taggedproteinRefoldingElution buffer:Binding bufferwith a higher concentrationof imidazoleGeneral purification of histidine-tagged proteinsBinding buffer(includ
10、ing 20 to40 mM imidazoleand 8 M ureaor 6 M guanidinehydrochloride)Binding buffer(including 20 to40 mM imidazoleand 8 M ureaor 6 M guanidinehydrochloride)Elution buffer:Binding bufferwith a higher concentrationof imidazoletagged proteincell proteindenatured tagged proteinFig 2.General purification wo
11、rkflow for histidine-tagged proteins(assumes use of Ni2+-charged affinity media,but other metal-ion-charged media follow the same workflow).11?1?1?+/-?+?/-?+/-+1+?-+2+?+/-+/-?-+3+=?-=?1?2?3?12?3?8?3?DNA?2?2?100%?Inner membranePeptidoglycanOuter membraneLipopolysaccharide70 70 70 210 Cytoplasm2000 pr
12、oteins100 proteinsPeriplasm10 proteinsMedium13?1?GST?4-10?GST?3?3?TEV?-?GST?GST?PreScissionTM Protease?4oC?GST?5?GST?GST?GE Healthcare?GST?GE Healthcare?A?-SepharoseTM 4B?A?IgG Sepharose Fast Flow?HiTrapTM Streptavidin HP?Strepsavidin Sepharose High Performance?14?96?His MultiTrap HP?His MultiTrap F
13、F?GST MultiTrap 4B?GSTMultiTrap FF?3?5?4?4?0-10mg/l?4?2.5mg/l?4?12.5?g50?g1mg10mg50mg?5ml20ml400ml4l20l?0.5ml1ml20ml200ml1000ml?3?GST?5?Cell wall disruptionRecover clarifiedsampleCell lysisHarvest inclusionbodiesInsoluble incytoplasmPeriplasmicspaceSoluble incytoplasmCell debris removalRecover clari
14、fiedsampleCulturemediumRecoversupernatantIntracellularexpressionExtracellularexpressionPurificationCell removalCell removalSolubilization15?5?5?Manton-Gaulin?French Press?2?1?0.2mg/ml?37oC?15?5?3?DNase?pH?16?DNA?3?8?HisTrapTM FF?His GraviTrapTM?His MultiTrap?3?GST MultiTrap?GST?5?GraviTrap?MultiTrap
15、?pH?/?17?1?pH?pH 2-pH9?pH?0.5M?0-2M?0-2M(NH4)2SO4?10oC?4oC?40oC?22oC?280nm?SDS-PAGE?-?190-800nm?HisTrap FF?His GraviTrap?His MultiTrap?3?GSTMultiTrap?GST?5?5?m?6?10000g?15min?40000-50000g?30min?10?15min?PVDF?6?18?6?1?m90?m?0.45?m30?34?m0.22?m3?10?15?m?“?”-?9?/?HIC?pH?pH?5000?100mM?100mM?SDS-?SDS-PAG
16、E?19?GST?3?5?280nm?GST?Lowry?BCATM?Bradford?DNA?SDS-PAGE?600nm?1?5-10l?8?SDS-PAGE?PVDF?HybondTM-C?Hybond-P?GST?SDS-PAGE?20?pH?7?7?SDS-PAGE?N?C?pH?N?21?GST?96?His GraviTrap?1ml?His GraviTrap?40mg?HiTrap?GST?HisTrap?GSTrapTM?HisTrap?ml?40mg?HiTrap?KTAdesignTM?HiTrap?96?GST?96?MultiTrap?1mg?HisMultiTra
17、p?0.5mg GST?GST MultiTrap?96?SpinTrap?96?His SpinTrap?750?g?GST SpinTrap?400?g?GST?22?KTAdesign?KTAdesign?KTAprimeTM plus?5?GST?HiTrap?KTAprime?5?KTAprime plus?6?KTAexplorer?KTAexplorerTM?6?8?KTAprime plus?KTAexplorer?23KTAexplorer?KTATM 3D plus Kit?KTATM 3D plus Kit?KTAexplorer 100?6?4?AC?GF?IEX?50
18、mg?90%?KTAxpressTM?7?KTAxpress?1?12?48?4?50mg?95%?7?KTAxpress?KTAdesign?KTAdesign?8?3?4?GST?5?24?8?KTAdesign?UNICORNTM?pH?cGMP?KTAdesign?KTAKTAKTAKTAKTAKTAKTAprimepuri?erexplorerxpresspilotcross?owproessplus?8?KTAdesign?rero l pxeATKsu l pemi rpATKKTApurifierKTApilotKTAxpressssecorpATKwo l f ssorcAT
19、K25?Ni2+?Ni Sepharose High Performance?HP?Ni sepharose 6 Fast Flow?FF?IMAC?8?1?9?8?4?26?9?Ni2+?NativeconditionsBinding buffer(including 20 to40 mM imidazole)Binding buffer(including 20 to40 mM imidazole)DenaturingconditionsCell lysisBinding toaffinity mediaWashElutePure denaturedtagged proteinPure t
20、aggedproteinPure taggedproteinRefoldingElution buffer:Binding bufferwith a higher concentrationof imidazoleGeneral purification of histidine-tagged proteinsBinding buffer(including 20 to40 mM imidazoleand 8 M ureaor 6 M guanidinehydrochloride)Binding buffer(including 20 to40 mM imidazoleand 8 M urea
21、or 6 M guanidinehydrochloride)Elution buffer:Binding bufferwith a higher concentrationof imidazoletagged proteincell proteindenatured tagged protein27?Ni2+?GE Healthcare?Ni Sepharose HighPerformance?Ni Sepharose 6 Fast Flow?IMAC Sepharose High Performance?IMAC Sepharose 6 Fast Flow,?Ni Sepharose Hig
22、h Performance?Ni Sepharose 6 Fast Flow?Ni2+?6M?8M?1?96?Ni Sepharose High Performance?Ni Sepharose 6 Fast Flow?Ni Sepharose High Performance?Ni Sepharose 6 Fast Flow?GE Healthcare?IMAC?1ml?5ml?HiTrap IMAC HP?HiTrap IMAC FF?20ml?HiPrep TM IMAC FF 16/10?IMAC Sepharose High Performance?IMAC Sepharose 6
23、Fast Flow?1?Ni2+?Zn2+?Co2+?4?Ni Sepharose High Performance?Ni Sepharose 6 Fast Flow?Ni2+?GE Healthcare?11-0008-86?IMAC Sepharose High Performance?IMAC Sepharose 6 Fast Flow?IMAC?28?Ni?0.22m?0.45m?2M?pH?pH 7?8?0.5M?1M?Tris-HCl?EDTA?280nm?8M?6M?8?SDS-PAGE?SDS-PAGE?Ni Sepharose?IMAC Sepharose?20-40mM?N
24、iSepharose High Performance?Ni Sepharose 6 Fast Flow?GE Healthcare?11-0008-86?4?29?280nm?HiTrap Desalting,PD-10 Desalting?HiPrep 26/10 Desalting?pH?2.5-7.5?pH?pH4?Ni2+?pH?EGTA?EDTA?HiTrap Desalting,PD-10 Desalting or HiPrep 26/10 desalting?4OC?7000-8000g?10min?1000-1500g?1.30min?2.?5-10ml?3.?4?4a.?0
25、.2mg/ml?20ug/ml?DNase?1mM MgCl2?1mM PefablocTM SC?PMSF?4oC?30min?4b.?10min?French press?5?HisTrap FF?30?5.?pH?pH?0.45m?0.22m?HisTrap FF?His GraviTrap?His MultiTrap HP?His MultiTrap FF?20mM pH 7.4?0.5M?0.5M?pH?7-8?HiTrap Desalting,PD-10 Desalting?HiPprep 26/10 Desalting?20-40 mM?6M?8M?8?31?10?Ni?10?N
26、i Sepharose?9?Ni Sepharose High Performance?Ni Sepharose 6 Fast Flow?72?9?Ni SepharoseHighPerformance?25ml100ml?40mg/ml?+?+?/?-?-KTAdesign?+Purification using precharged mediaBuffersincluded?Buffersincluded?His MultiTrap FFHis MultiTrap HPNi Sepharose 6 Fast FlowHis SpinTrapYesNoYesNoHis GraviTrap K
27、itHisTrap FF crude KitHisTrap FF crudeHisTrap FFHis GraviTrapSelection GuideNi Sepharose products PrechargedWill you usean automatedpurification systemsuch asKTAdesign?Whatare yourrequirements?NoYes96-wellplateBatchSpincolumnGravity-flowcolumnSyringePrechargedNi SepharosemediaContains Ni Sepharose H
28、igh Performance.Contains Ni Sepharose 6 Fast Flow.32?HisTrap HPHis SpinTrapHis MultiTrapHP?1ml5ml100?l9 6?40mg?200mg?0.75mg?1mg?-+?-+-?/?-?+?-KTAdesign?+-?9?Prepackedcolumns?Ni SepharoseHigh PerformanceHisTrap HPNi Sepharose 6 Fast FlowYesNoClarifiedor unclarifiedsampleHisPrep FF 16/10HisTrap FF cru
29、deHisTrap FFNoYesPrepackedcolumns?HighresolutionScalability/high flowratesClarified200 mgUnclarified100 mgHiPrep IMAC FF 16/10Cu2+?Zn2+?pH?HiTrap IMAC FF 5ml?HiPrep IMAC FF 16/10 20ml?90%?43?75%?12?90%?Zn2+?HiPrep IMAC FF 16/10?3%?rBCA?Zn2+?HiPrep 26/10 Desalting?43?A?HiTrap IMAC 1ml?B?HiTrap IMAC 5
30、ml?C?HiPrep IMAC FF16/10 20ml?2.4ml?12ml?49ml?12.5?62.4?255mg r-BCA?74%?D?SDS-PAGE?ExcelGel Gradient 8-18?1%?PhastGel Blue R?94?12?IMAC Sepharsoe 6 Fast Flow?r-BCA?r-BCA?HiTrap IMACFF 1mlHiTrap IMACFF 5mlHiPrep IMACFF16/10(20ml)?mg?12.5-62.4-255-?mg?-11.7-56.1-235?mg?-94%-90%-92%?r-BCA?93%-84%-90%?1
31、3?SDS-PAGE?13?His?ELISA?His?ECL?SDS-PAGE?HRP?ECL?ECL?ECL AdvacneTM?SDS-PAGE?6SDS?0.35M Tris-HCl?10.28%?W/V?SDS?36%?V/V?0.6MDTT?5%?-?0.012%?w/v?pH6.8?0.5ml?-80oC?5-10l?2l 6 SDS?1.?90-100 oC?5?2.?SDS-?3.95?Coomassie Blue R Tablets?PlusOne Silver4.Staining Kit?SDS-PAGE?14?14?SDS-PAGE?%?5%7.5%10%12.5%15
32、%?5-15%5-20%10-20%?Mr103?36-20024-20014-20014-10014-60114-20010-20010-150?Western blot?Western blot?ECL?ECL plus?ECL Advance?His?/?5%?w/v?0.1%?v/v?Tween20?PBS?140mM?2.7mM?10mM?1.8 mM?pH7.3?0.1%?v/v?Tween 20?PBS?His?Ig?HRP?Whole Ab?NA931?SDS-PAGE?1.GE Healthcre?His?2.?Hybond ECL?ECL?Hybond P?ECL?ECL
33、plus?ECL advance?GE Healthcare?Hybond ECL?1.?50?200?/?2.?1-16?3.?4.?16?96?/?His?5-10l?50ml?1.?GE Healthcare Application Note19-1139-13?2.?1?3.?4.?20-30ml?/?5.?6.?20-30ml?/?10-60?7.?8.?/?GE1.Healthcare Application Note 19-1139-13?2.?1?3.?4.?20-30ml?/?5.?6.?20-30ml?/?10?60?7.?8.?9.?Western blotting?GE
34、 Healthcare ApplicationNote19-1139-13?14-0003-87?ECL?ECL plus?ECL Advance?-?GE Healthcare?His?Western blot?His?97?GST?rTEV?Invitrogen?6?Glu-Asn-Leu-Tyr-Phe-Gln?Gly?Gln?Gly?Glu?Tyr?Gln?Gly?N?6?Ni Sepahrose?6?6?rTEV?SDS-PAGE?SDS-PAGE?rTEV?29000?Carboxypeptidas A?C?94000?PreScission?9?98?KTAxpress?KTAx
35、press?KTAxpress?AcTEV?44?APC234?32500?30000?AC?DS?IEX?GF?KTAxpress?SDS-PAGE?SDS?7.5g?44?A?6?AcTEV?B?SDS-PAGE?99?His SpinTrap?His GraviTrap?0.22?0.45m?1?HisTrap FF Crude?/?DNase?Mg2+?DNaseI?5g/ml?Mg2+?1mM?10?15?4C?100?pH?8M?6M?SDS?2%TritonX-100?2%Tween 20?2%NP-40?2%cholate?1%CHAPS?1.5M?50%?20mM?-?1-3
36、mM DTT?DTE?5mM?5mM TCEP?10mM?8M?6M?30?TritonX-100?NP-40?Tween?280nm?4-6M?4-8M?30?pH?pH?4-8M?4-6M?0.2%?TritionX-100?/?pH?101?Western blot?His?/?/?/?SDS-PAGE?260nm?0.1?10mg/ml?25mM Tris-HCl?pH8.0?pH?4-6M?4-8M?20mM?8M?6M?pH7.4?7.6?500mM?5-40mM?SDS-PAGE?EDTA?20?102?/?2%?TritonX-100?2%?Tween20?50%?/?pH?4
37、-8M?4-6M?HisTrap?2-3?KTAdesign?0.5MPa?5bar?0.3MPa?0.2MPa?MultiTrap?500l?MultirTrap?103?1.?45?Mycobacterium bovis?G?(His)6-PknG?45mM?Ni sepharose HighPerformance?45A?45B?SDS-PAGE?45mM?104?45?A?B?45mM?(His)6-PknG?2?26ml?0.45m?2ml NiSepharose High Perforamnce?XK16/20?KTApurifier?50%?100%?C?12%SDS-PAGE?
38、(His)6-PknG?+?45mM?-?1052?His SpinTrap?His SpinTrap?APB7-(His)6?28000?5?50?100?200mM?His SpinTrap?500mM?5mM?46?3?50mM?46?4?100mM?46?5?200mM?46?6?20?40mM?46?APB7?SDS-PAGE?ExcelGel Gradient 8-18?3?4?5?6?106?pH?Ni2+?Ca2+?Co2+?Cu2+?Fe3+?Zn2+?GE Healthcare?IMAC?1ml?5mlHiTrap IMAC HP?HiTrap IMAC FF?20ml H
39、iPrep IMAC FF 16/10?IMAC SepharoseHigh Performance?IMAC Sepharose 6 Fast Flow?3?Ni2+?Co2+?Cu2+?Zn2+?Cu2+?Cu2+?Zn2+?Cu2+?Zn2+?Fe3+?Ca2+?Fe3+?pH?3?Fe3+?Fe3+?50mM EDTA?1?HiTrap IMAC FF?Cu2+?Zn2+?Ni2+?BL-21?6?APB7?28000?Cu2+?Zn2+?Ni2+?HiTrap IMAC FF 1ml?IMAC Sepharose 6 Fast Flow?47?A?C?20mM?Cu2+?Zn2+?N
40、i2+?10mM?Cu2+?Zn2+?107?20mM?5mM?Ni2+?20mM?47?SDS-PAGE?HiTrap IMAC FF 1ml?IMAC sepharose 6 FastFlow?APB7?Cu2+?Zn2+?Ni2+?A?20mM?B?10mM?C?5mM?SDS-PAGE?IMAC Sepharose?IMAC?20-40mM?280nm?Hitrap Desalting?PD-10 Desalting?HiPrep 26/10 Desalting?1082?HiTrap IMAC HP?Cu2+?Zn2+?Co2+?Ni2+?IMACSepharose 6 High P
41、erformance?HiTrap IMAC HP 1ml?Cu2+?Zn2+?Co2+?Ni2+?APB7?28000?ABP7?Co2+?Ni2+?Cu2+?Zn2+?48?48?BL-21?APB7?A?Cu2+?B?Zn2+?C?Co2+?D?Ni2+?HiTrap IMAC HP 1ml?1ml?SDS-PAGE?ExcelGel SDS Gradient 8-18?E?109?7?1?His?10?(His)10-trx-P450?N?10?HisTrap FF?HiLoad 16/60 Superdex 200 pg?49B?5?6?2?7?8?3?4?N?9?1?(His)10
42、-trx-P450?49?A?(His)10-P450?B?SDS-PAGE?1102?KTAxpress?100ml?HisTrap FF crude 1ml?AC?HiPrep 26/10?DS?Mono QTM 5/50 GL?IEX?AC-DS-IEX?SDS-PAGE?50?A?ACDSIEX?IEX?1?2?9.4mg?B?IEX?SDS-PAGE?111?GST?S-?GST?1988?GST?GST?GE Healthcare?GST?GST?pGEX?GST?GST?GST?GST?GST?pGEX?Schistosoma japonicum?GST?26Kd?pGEX?S.
43、japonicum?GST?N?GST?GE Healthcare?Sepharose High Performance?HP?Sepharose 6 Fast Flow?FF?Sepharose 4B?Glutathione Sepharose?96?HiTrap?HiPrep?25ml?500ml?GST?pGEX?GST?GST?GST?18-1157-58?GST?112pGEX?GST?pGEX?tac?IPTG?pGEX?lacIq?tac?IPTG?15?15?pGEX?pGEX-6P-1?pGEX-6P-2?pGEX-6P-3pGEX-4T-1?pGEX-4T-2?pGEX-4
44、T-3pGEX-5X-1?pGEX-5X-2?pGEX-5X-3pGEX-2TK?PreScission Protease?Xa?EcoRI?7?pGEX-6P-1?pGEX-4T-1?pGEX-5X-1?gt11?cDNA?pGEX-2TK?cAMP?-32PATP?7?pGEX?DNA?GEHealthcare?http:/ PreScission?GST?GST?113?pGEX?Lon?OmpT?DegP?HtpR?BL21?pGEX?SGS-PAGE?BL21?OmpT?Lon?GST?BL21?JM105?pGEX?recA1?DNA?DNA?DNA?2Kb?PCR?GST Mul
45、tiTrap 4B?96?GST?600l?2-3ml?Glutathione Sepharose?GST SpinTrap?12ml?8?600nm?114?GST?Sepharose?51?Glutathione Sepharose?GST?Glutathione Sepharose?GST?GST?51?Glutathione Sepharose?SH?Sepharose?Sepharose?GST?115?GST?GST?16?Glutathione Sepharose?2?16?GST?GlutathioneSepharoseHighPerformanceGlutathioneSep
46、harose 4Fast FlowGlutathioneSepharose 4 BGlutathioneSepharose 4 BGST MultiTrapFFGST MultiTrap4BGST SpinTrap?GSTrap HP?/?25ml100ml25ml100ml500ml10ml100ml?300ml2ml50?l50?l50?l1ml5ml?/?10mg?rGST?10mg?rGST?5mg?rGST?10mg?GST?0.5mg(rGST)?0.5mg(rGST)?0.4mg?rGST?10mg?50mg?rGST?2?GlutathioneSepharose 4 B?Glu
47、tathioneSepharose 4 FastFlow?96?GlutathioneSepharose 4 B?96?50?GlutathioneSepharose 4 B?GlutathioneSepharose HighPerformance?116?GSTrap FFGSTrap4BGSTPrepTMFF16/10pGEX?GST?GST?HRP?GST?GST96?ECL GST?/?1ml5ml1ml5ml20ml5?25?g?0.5ml75ul5?HRP?GST?GST?1000?3000cm2?/?10mg?50mg?rGST?5mg25mg(?GST)?200mg?rGST?
48、N/A50?1:5000?N/AECL Plus?A?ECL Plus?B?GlutathioneSepharose 4 FastFlow?GlutathioneSepharose 4 B?GlutathioneSepharose 4 FastFlow?BL21?GST?HRP?GST?ECL?GST96?tac?PreScission?Xa?GST?GST?GST?GST?GST?GST?HRP?GST?117?PreScissionProtease?XaHiTrapBenzamidineFF?HighSub?BenzamidineSepharose4 Fast Flow?High Sub?
49、/?500?500?400?1ml5ml25ml96?/?90%?100?g GST?1mM EDTA?1mMDTT?150mM?,50mM Tris-?(pH7.0)?5C?16?90%?100?g?GST?22C1PBS?16?90%?100?g?GST?1mM?100mM?50mM Tris-?(pH8.0)?22C?16?35mg?175mg?35mg?/?500?l?PreScissionProtease?Xa?BenzamidineSepharose4Fast Flow?High Sub?V?Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro?Gln?Gly?GST?P
50、reScssion?pGEX-6P?GST?pGEX-T?GST?Ile-Glu-Gly-Arg?Xa?pGEX-X?GST?Xa?Xa?GST MultiTrap?118?GST?1mg/l?10mg/l?17?2.5mg/l?17?GlutathioneSepharose?1PBS1?50mg20l1l10ml100ml10ml10mg4l200ml2ml20ml2ml1mg400ml20ml200?l2ml200?l50?g20ml1ml10?l100?l10?l?0.45m?DNase?RNase?GST?Glutathione Sepharose?GST?HiTrap Desalti
