UNICORN Ver.4.1版 - GEヘルスケアバイオサイエンス
Contents
1. Hun Run 13 14 11 2 1 pH IE pH 65 66 RESOURCE S 1 ml Buffer CIEX 3 0 7 5 4 mMmin pH 7 6 5 4 5 CV P 950 12. Injection technique Manual 143 0 1 1 1 Cross sectional diagram 1 Upper outer end piece 2 Protective jacket 3 Inner end Diece 4 Bottom outer end Diece 5 Inner end piece 6 Movahle seal 7 Glass tube 8 O ring 11 3x2 4 mm 9 O ring 27 1x1 6 mm 10 11 12 13 14 19 16 17 18 Milli Q Union 1 16 female M6 male PEEK 18 1112 57 6 Fingertight Connector 1 16 18 1112 48 10 PEEK
3. tr 2 lt 3 A TN OFF
4. 1 1 Wizard Method EditOr Flim Ll METIHN mi tmmi File NewW New Method Wizard OK Teri haingd E cfr mrt Fr ramurm ee i 2 nl mim hm rslmam rm 42 Technique Affinity Anion Exchange Cation Exchange Chromatofocusing CIP HIC RPC Size Exclusion
5. Column 20 2 Column Position BufferPrep ON OFF Next gt Wavelength UV1 Pump A B Inlet Buffer Valve A Inlet Start Concentration B Equilibration Volume 1 2 3 9 Variables A1 B1 A1
6. Manual Flowpath InjectionValve Load Execute 10 86 12 1 28 8S1 82 P 950 1 5 Frac 950 Frac 900 Frac 900 Frac 900 gt P 87 lt Frac 950 gt P 950 950
7. Manual Flowpath BufferValveA1 At11 Execute A11 ml 145 0 1 2 Method EditOor 2 Injection technique System Pump Direct Loading Injectlon Volume 2 5ml eXDlorer 100 2 9mm 8 3ml 1 0mm 1ml
8. P 950 P 950 3 2 0 6 43 2 Load Shaded area covers the components located Inside the valve door linefilter Column MS Selection Valve Injection V2 Pump EP Valve V1 COND UV O gt gt J a A2 B1 raction a collector optional Buffer Outlet Valve V4 2 4js Column EY Selection W3 F3 Valve V3 3 5 Figertight connector 1 16 18 1112 55 PEEK 1 0mmX50cm Figertight connector 1 16 18 1112 55 Union 5716 female
9. Next gt 47 8 Elution mi Elution Technigue lsocratic lsocratic with Delayed Fractionation 7 Linear Gradient Segmented Gradient Segmented Gradient HIC Linear Gradient Target Concentratin9oB B Gradient Length CV B9 CV Wash Out Unbound Sample 2 CV Clean after Elution at 100 B 5 CV
10. UV 17 1 127 UV explorer 10S pH explorer 100 0 5MPa gt 10mMmin 2 1 UNICORN 1 ON ON 3 Windows Begin Logon Ctr Alt Delete
11. 5 Load S 1 LOAD 5 Load Manual Flowpath InjectionValve Inject Execute
12. Undo zoom Reset ZOOm 116 Point LisSt Delete Delete all Draw baseline 2 2 Draw straight to next Doint OK Edited Baseline OK
13. HiPrep 0 15MPa AKTA 0 35 MPa HiPrep 0 15 MPa FR 902 0 2 MPa UNICORN 0 5 MPa 6 6 1 1 2 Manual Pump Pumpwash Explorer A11 ON ON ON Execute
14. OFF ON 10ml Injection Volume Next gt Frac Frac 950 gt Frac 950 Rack 18 amp 30mm Tubes Flowthrough Fractionation Frac 950 Fraction Volume 50 ml Tube Type 30 mm Start at First tube lt Frac 900 gt Flowthrough Fractionation Frac 900 Fraction Volume 5 ml Next gt L a wt ul Elution Fractionation Frac 950 gt Fixed Volume Fractionation Frac 950 Fraction Volume 1 ml Tube Type 18 mm Start at First Tube lt Frac 900 gt Fixed Volume Fractionation Frac 900 Fraction Volume 1 ml Next gt 61 7 Peak Fractionation Peak
15. Slope 1 Slope X Time min 2 Operations Differentiate UV First Order OK ie a _ meiTl i fmdet onmidnmder mf Cam Hmn 3 IMarker EMH _ Wi 121 4 Slope XY slopeg 5 Slope XY 6 Slope Method Editor Peak_FracParameters 14 10 Export
16. A11 A2 B1 B2 Pump 1 A11 A2 B1 B2 2 A11 B1 30ml L Po ML Ut 3 4 ml 5 2 4 6 A2 B2 System Control Manual Flowpath PumpAInlet A2 ExeCute
17. 2 SpIlit Peak 1 2 Join left Join right Peak start Peak end 1 2
18. 0 1 2 1 1 2 0 45 um 8 A18 HIC System Pump Dilrect Loading B2 A B 100 B 096B A18 A18 Manual Flowpath BufferValveA1 A18 Execute A 30ml
19. 14 2 8 P 99 File Report Format 5 Globall All ChromatogramSs Print OK 14 4 14 4 Run Log 14 4 1 1 File Report 2 Generate ReDort TE TI ul gh 1 LIP PPT Ei Benu HmPd 3 4 Print Report Preview Customize Report 14 4
20. 11 2 RESOURCE HIC 1ml PHE ISO ETH Buffer A 0 1M Phosphate 2M pH7 2 B 0 1M Phosphate pH7 2 4 ml min CV P 950 1 0 1 ml 0 100 10 CV 100 5CV 280 nm 254 nm 214 nm Position 2 Position 3 Position 4 Frac 950 18 amp 30mm gt 1 50 ml 1ml Frac 900 A B
21. 8 1 3 A B C 12mm EL ITITITLIE Tll PTTTTTLTTTI 18nmm amp 30mm A 10 100ml min ml 18mm Yellow 18mm gt 15ml 30mm Red 30mm gt 50ml 8 1 4 DropSyncro Accumulator Accumulator B 10mlmin 10 ml 18mm Yell
22. Tu He Ire 0 Fi 7 PEEK HHECT LT 1 1 i 1 1 LE _ ee 1 1 a Ed i a hh a 1 WNW 1 Lock Knob 5 mm 41 9 ff 9 1 Wizard
23. 10ml 50ml 150ml 3 V2 V3 7 1 V5 7 8 P 950 0 1 50mlmin 1 0MPa FR 902 0 2MPa FR 904 0 4MPa 2 pH FR 902 FR 904
24. 2 3 118 14 6 8 14 1 2 2 Edit Peak Name Peak name OK 3 1 2 4 OK 5 6 Properties Chromatogram Layout Curve Style and Color 7 Peak label Peak Name 8 OK
25. Reset ZOOm 13 2 3 13 2 4 LogDbook 94 View 1 Panes Flow Scheme amp View 1 Panes LO9gDOOkK amp
26. amp 4 XY 5 marker 104 105 14 2 1 1 1 Add Text 2 abDC 3 abC 4 text gt text gt
27. 7 8 P 950 5 ml min Manual Pump SampleFlow 5mlmin Execute 9 0 11 System Control End P 950 8 Frac 900 P 40 8 2 Frac 900
28. FE 4 Page DOown Page UD 5 6 Reset ZOOm 14 2 7 1 File Opentocompare Chromatograms Hm Lh rem ml rn lo CNet Lmao et i a i a Ed imrrisirnr mrra Le ed Of _ HE 2 Chromatogram selection 3 Browse 4 IeX IEX lex jex jex iex 2IeX 4 iex
29. OK 2 Marker 3 COnd 4 Cond UV 280nm 119 14 8 120 1
30. Notes Gradient BufferPrep BufferPrep 11 Columns Reference CurVeS System Control Evaluation DrocedureSs Method information Settings System settings Calibration System Control Syst
31. Evaluation Microsoft Word Paste 1 9 2 Edit Copy to clipboard 3 Windows NT Paint Paste Bm Muma mmTem Cecuam i Ciaracser Mag eee 3 Pe a Ammanve Tous Iamal HN PRook Viewm PPmrahiors IFomnmml Cock o Meroso WWes r brempia pe ae ems A ee me a Ma Ro EE onerd Ph Mir Ectange nf a We HT Ek 14 12 EvaluatlOn 1 Operations Create Curve Er Et eh Fs Chiun El LihFumm erie r
32. 1 pH KCI 2 3 pH7 0 4 System Calibrate Monitor pH Reference Value1 6 86 25C Er PPT mmmrmmmrmm imi Fan mitt E Si Eh fh EE EL 135 5 Measured value pH Read Value1 9 Main menu 4 6 pH4 01 25C pH9 18 19 1 7 Reference Value2 4 01 25C pH9 18 8 Measured value pH RHead Value2
33. 10S 2um 2M NaOH 18 1120 94 100 18 1027 11 Frac 900 95 10 18mm 175 12mm 100 40 30mm 3 Frac 950 120 18 amp 30mm A 240 12mm B 45 30mm D 96 4 amp 30mm 8 C 4
34. VVizard Sample Injection Technique Sample Injection Technique 0 2ml Manual 0 1 2ml SamplePump Loop Filling 1 150ml SamplePump Loop Filling 20L SamplePump Direct Loading gt 10ml SystemPump Direct Loading BufferPrep 20 1 1 10 50 150ml 10ml 18 1113 81 50ml 18 1113 82 150ml 18 1023 85
35. 0ml Upper Chamber PEEK 150 ml 0 O 0ml Bottom Chamber Bottom Chamber PEEK Upper Chamber 6 Bottom Chamber 2
36. A11 B 0 CV 3 5CV Next gt 43 44 4 Injectlion Technidue Manual Sample Pump Direct Loading P 950 Sample Pump Loop Filling P 950 System Pump Direct Loading Injection Technique Manual Empty Loop with 3 1
37. Method Editor File New New Method Wizard OK Technique Cation Exchange Column RESOURCE S 1 ml Column Position Position2 BufferPrebp OFF Next gt 3 Wavelength UV1 1 280 nm 2 3 Variables Pump Inlet Pump A Inlet A1 Pump B Inlet B1 Buffer Valve A Inlet A11 Start Concentration B 0 B Equilibration Volume 5CV CV A 3 5CV Next gt 4 Fr lr mmm FE Le hi EE ra EN Lam ph TFT ml FT Hre Eos Laon WER roi Injection technique Fill Loop with Empty Loop With Sample Inlet Sample Pump Loop Filling P 950 0 2 ml 85ul 0 2 ml
38. 3 Properties 4 Chromatogram Layout Apply to all chromatograms 0K 14 2 9 102 1 File Opentocompare Curves Open Curves to Compare eho EEWII TI 1 lmm WENT EE LET lrmm ll hm CEE CHL lm EE OT IFFI mm wlmi HI mi ima CET MOT ND Le m 2 Chromatogram selection 3 Browse 4 UV 3 UV UV1_280 UV1 280 5 Searchnh 6 7 Store in new chromatogram
39. 2 3 S1 S 8 Rinse Sample Loop 0 5 ml 1 Next gt Frac Frac 950 gt Frac 950 Rack Flowthrough Fractionation Fraction Volume Tube Type Start at Frac 900 gt Flowthrough Fractionation Fraction Volume 18 amp 30mm Tubes Frac 950 5 ml 30 mm First tube Frac 900 5 ml Next gt Elution Fractionation Frac 950 gt Fixed Volume Fractionation 0 Fraction Volume Tube Type Start at Frac 900 gt Frac 95 1 ml 18 mm First Tube Fixed Volume Fractionation 0 Fraction Volume Frac 90 1 ml Next gt Peak Fractionation Peak Fractionation OFF Next gt 8 Elution
40. 0 2 1 2 Method Editor Edit Column List Column List Cacia Fm Da il TL Fn a Valve Replace Typical peak width at base Average time Peak fractionation Min Width Save AS SaVe as dloODal Save as glODal Save AS 14 0 3 0 3 1 Text Instruction
41. AKTAexplorer 10S 100 P 903 AKTAexplorer 10S 2 0 001 10mmin 25MPa P 901 AKTAexplorer 100 2 0 01 100ml min 10MPa UV 900 190 700nm UV Vis 3 explorer 10S 10mm 8ul explorer 100 2mm 2ul pH C 900 pH AKTAexplorer 10S 100 Flow Direction Valve V7 Outlet Valve V4 only in AKTAexplorer 100 Buffer Valve V6 Column Selection Valve V2 Injection Valve V1 Box 900 um _ Monitor pH C 900 D Monitor UV 900 a Sample Valve V5 only in AKTAexplorer 100 and AKTAexplorer 10S Column Selection Valve V3 Pump P 900 P 901 alt P 903 Shaded area covers the components located inside the valve door G5 Flow _ On
42. System Control 4 Start protocol Next Systems 1 Settings Specials FracParameters DelayVol lt Frac 950 gt Frac 950 RHack Fraction order AKTAexplorer 10S Last tube Define Rack pH Cell pH 0 25mm 105 uil 120 NG 0 5mm 301 ul 423 pl AKTAexplorer 100 pH Cell pH 0 5mm 297 ul 419 Hl A 0 75mm 624 pl 773 ul pn 1 0mm 1087 Hl 1267 ul F 2 ET pH FF Le pH 18 2 pH P 129 pH
43. 001 Resull 11 UNICORN Main Menu Methods 10 3 RESOURCE S 1 ml 4 ml min 5 CV 100ml 0 50 B 5CV 50 1009 B 0CV 100 B 3 CV 280 nm 254 nm 214 nm Position 2 Frac 950 18 amp 30mm Tubes gt 50 ml 30mm 1 ml 18mm Frac 900 1 Method Editor File New New Method Wizard OK 2 Technique Cation Exchange Column RESOURCE S 1 ml Column Posi
44. 3 5CV Next gt 4 ErE Il rr ee Ti Pr Le Fn FE LS hi EEE mn TE a i rd Injection Technique Sample Pump Loop Filling P 950 Fill Loop with 2 ml 85M Empty Loop with 2 ml 2 3 Sample Inlet S1 S G Rinse Sample Loop 0 5 ml 1 Next gt Frac Frac 950 gt Flowthrough Fractionation Frac 950 Fraction Volume 5 ml Tube Type 30 mm Start at First tube Frac 900 gt Flowthrough Fractionation Frac 900 Fraction Volume 5 ml Next gt Elution Fractionation Frac 950 gt F
45. 7 1 1 3 4 2 18 1127 66 3 P 950 0 7mm 18 1801 42 P 950 3 3 Load Shaded area COVers the Wa I i 1 4 On Ilinefilter 5 ICN 6 S1
46. TEL 03 5331 9330 Ne Ce FF 9 00 17 30 9 TEL 03 5331 9315 FAX 03 5331 9349 TEL 06 6305 4707 FAX 06 6305 3599 TEL 03 5331 9336 2 9 4 FAX 03 5331 9370 e mail Tech JP ge com O amp A
47. 1 2 Next gt lt Frac 950 gt em mr jm si Frac 950 Rack 18 amp 30mm Tubes 12mm Tubes 30mm Tubes MicroPlates amp 30mm Tubes Flowthrough Fractionation Frac 950 Frac 950 Outlet Valve F3 3 F3 OFF 3 Fraction Volume ml Tube Type 18mm 30mm Start at NextTube FirstTube 1 lt Frac 900 gt Flowthrough Fractionation Frac 900 Frac 900 Outlet Valve 3 F3 OFF
48. Manual Frac FractionationStop 900 Execute Manual Flowpath OutletValve F3 Execute 3 END 2 209 2 15 1 3 System Control 1 Manual Pump PumpWeash Explorer A11 ON ON ON Execute 15 1 Manual Pump SystemWash 40ml explorer 10S 75ml explorer 100 Execute 6 15 3 i 15 1 S1 5 5 2 3 Manual Flowpath SampleValve Execute 4 Manual Pump SampleFlow 2mlmin ExeCcute
49. 1 2 w LOAD Union Luer female 1 16 male PEEK 18 1112 51 2 3 20 1 BufferPrep
50. 3 P 950 P 950 2 1 G5 3 Inject Shaded area covers the components located Inside the valve door On Iinefilter Column Mixer Selection Valve 2 Injection V2 Pump Pump Valve V1 SS COND UV O gt J U0 9 Fract A2 B1 C raction a collector optional Buffer Outlet Valve V6 Valve V4 a LA 4 RB JUUUUUUU Column A11 A12 A13 A14 A15 A16 A17 A18 Valve V3 W3 F3 1 9 7 5 2 Union 5716 female HPLC male 18 1142 08 Teflon 1 6mmX50cm Tubing connector for 18 o d tubing 18 1121 17 with ferrule for 1 8 o 0 tubing 18 1121 18 Tubing connector for 1 8 o d tubing 18 1121 17
51. 20 2 3 4 A B 16 3 2M NaOH explorer 10S 18 1120 94 100 18 1027 11 16
52. Pressure 17 18 8 A A2 Manual Pump Gradient Target0 B Execute Manual Flowpath PumpBInlet A2 Execute A11 A2 B1 B2 0 05MPa 9 End 2 209 4 1 1 System Control 2 Manual Pump PumpWeash Explorer A11 0ON ON ON Execute 5 PumpWash
53. 8 CurVe ODtIOnS Overlay Stack 9 OK 10 Stack Shift Curves by Offset Offset 11 Y Shift Curves by Offset rmt i ri LE i i TT eS ie Th rr iT ll Ir HH Im lmmm HT LL Em 103 1 1 2 Properties 2 File Open Curve 3 Chromatogram Layout mimrm mm LE ED Y AxIS
54. Method Editor UNTITLED File Edt Block Wiew Hel Run Setub Text Mode 32 5 SampleFLow GamDpleFlOw Instruction Box ParameterSs Change SampleFlOow 10 5 Parameters Var VariaDle name difinitlOn SampleFlow OK Variables Variables gt P 34 8 0 15 SampleFlow 10 ml min 0 15 SampleFlow 5 ml min Eh EE 3m Pihaem je Ps 6 SampleFlow 0 00 ml min BreakPoint 1 15 2 15 Change BreakPoint Var
55. Waste W1 4 2 1 Manual Pump SystemWash 40ml explorer 10S 75ml explorer 100 Execute 5 SystemWash Load Position1 Bypass UV pH W3 4 3 P 950 7 24 H5 H12 H9 H8 V2 pH COND UV G3 G4 G1 G2 H7 V1 UN 2 B1 Ag Wi LAI IL_ IN 5
56. 10ml min P 950 1 8 0 75mm Pump P 950 Fingertight connector 1 16 Outlet connection Union 5 16 female HPLC male Tubing connector for 1 8 o d tubing with ferrule for 1 8 o d tubing TE 122 Inlet connection 30 P32 gt Purge method 1 S7 969 2 S6 3 S1 5 4 S8 A 5
57. ON OFF Enter lt VariaDleS gt Wavelength 2 3 Wash Out Unbound Sample Clean after Elution at 100 B Reequilibration Variables Show detallsS Frac 950 gt Variables Frac 950 Rack Wizard Fraction order Serpentine row 4 9 10 File SaveAs Method Name Techniqdue Next gt
58. Teflon tubing i d 1 6 mm o d 1 8 3 m 18 1121 16 Inlet filter assembly 1 18 1113 15 Inlet filter set for 10 ml 10 18 1113 16 PEEK tubing i d 0 25 mm o d 1 16 Blue 2 m 18 1120 95 PEEK tubing i d 0 50 mm o d 1 16 Orange 2m 18 1113 68 PEEK tubing i d 0 75 mm o d 1 16 Green 2 m 18 1112 53 PEEK tubing i d 1 0 mm o d 1 16 Gray 2 m 18 1115 83 Tefzel tubing i d 0 25 mm o d 1 16 2 m 18 1121 36 Tefzel tubing i d 0 50 mm o d 1 16 2 m 18 1120 96 Tefzel tubing i d 0 75 mm o d 1 16 2 m 18 1119 74 Tubing kit i d 0 25 mm for Purifier 1 18 1123 34 Tubing kit i d 0 50 mm for Purifier 1 18 1123 33 Tubing kit i d 0 25 mm for Explorer 10S 1 18 1122 12 Tubing kit i d 0 50 mm for Explorer 10S 1 18 1123 21 1 Manual Flowpath ColumnPosition Execute Manual Pump Flow 0 5mmin Execute 2 3 END
59. 1 Text InstructlOnS5 2 WindoOwWsS LlnrmmF mgeHnwyg FT rl FPPM FE nsh bo HE rd La He 3 Text Instruction box QOK ml le i 4 Haag li a i Il Bach PLCWm_THPEFGHCSLECTIEh Hamhie ire F Jl HH FE LE ci ELE ioT EN 1 1 EL ir 1 F di Ee eh Il Fn 1 im ET mi Fi 148 1 Text 2 3 Instruction box Breakpoint Instructions Parameters Insert 20 3 1 2 1 Text 2
60. LT Imm ED Im 1 UW em bs lm s I R ie i ml mm FF mm wg wm 1 a rf Ri mw Hh 5 3 6 Fixed 4 7 All with this unit 5 OK 9 6 Select gt gt II 8 6 14 8 10 7 3 6 1 8 OK marker XY 2 Ei 3
61. 3 5CV 9 Next gt 4 i pT hE EE TE La Fr Egy Lcrh mh MF mm Fa LL 1 Hrrm Sarnia La ES ni Injection technique Sample Pump Loop Filling P 950 Fill Loop with 0 2 ml 85 Empty Loop With 0 2 ml 2 3 Sample Inlet S1 S 8 Rinse Sample Loop 0 5 ml 1 Next gt Frac Frac 950 gt Frac 950 Rack 18 amp 30mm Tubes Flowthrough Fractionation Frac 950 Fraction Volume 5 ml Tube Type 30 mm Start at First tube Frac 900 gt Flowthrough Fractionation Frac 900 Fraction Volume 5 ml Next gt Elution Fractionation Frac 950 gt Fixed Volume Fractionation
62. 4 5 OK X X Time min Volume ml 1 CurveSs PropertieS 2 X AxIS Ci i 3 Time min Volume ml 4 Total Window Total Window OK 93 Undo Zoom
63. ASCII Lotus1 2 3 V WKS 1 EValuation 2 File Export Curves TN Ta EhL CL TE mm Fr mi ET LT mm I HEFAREPE T IF Gm Ta mm Fa HFTHEPS LT fe ET 1 Cd tl TE TIEFE Cor EE Co ei FT a TF HEHEFS 1 fi NE i I FEE PE 1 EL Fara br tmrig HETUFE 1 mm br re ami gt Pe Harimhrm mhmmipm Em me le 3 Source chromatogram 4 Cut curve Expot 5 Reduce number of samples by factor Expot factOr 6 Select EXDOrt 7 Save In 3 1 2 Floppy A DOS V 8 OK 122 14 11
64. Union M6 female 1 16 male PEEK 18 1112 58 8 O Tubing End fitting 19 2148 01 HiPreDp M6 Union 1 16 female M6 male PEEK 18 1112 57 6 0 5mm PEEK 100 0 75mm PEEK Fingertight Connector 1 16 18 1112 48 10 _ Fingertight Connector 1 16 10 18 1112 55 for 1 16 o d tubing PEEK Tubing connectoOr 10 18 1112 49 for 3 16 o d tubing PEEK Ferrules for 3 16 o d tubing PEEK 10 18 1112 48 UNion 1 16 female M6 male PEEK 6 18 1112 57 Union M6 female 1 16 male PEEK 8 18 1112 58 Union Luer female 1 16 male PEEK 2 18 1112 51 Stop plug 1 16 PEEK 5 18 1112 52 Stop plug 5 16 PEEK 5 18 1112 50 Union 1 16 female 1 16 female 1 18 3855 01 titanium Union Luer female M6 female 1 18 1027 12 Tubing connector M6 male 5 19 1993 50 SH 2
65. 2 File Open Curves Open CurveS lshbuma1 Fst Fm gglshbuFU HE en F ET 15 bis ETHIE enm F 11 0 i sii 11RKB si F WI7eUT 1 gitestD3 TCATEE Flies Fie Br TO 13 mumlFF Saep SOD Acc BITHR Fest Fle TAN E D3 marma FF Slimnii hmprml FE am F ML PL EL ET _Bemew I lma Ti ChhnmtalnmtmT 1 mm ceew He 3 Result 4 Select Selected CurVeS 5 RermOVe 6 OK 1 Operation Subtract 2 3 OK 14 9 Slope Slope Slope
66. ee a Pump PBPO IN 3 a 7 3 5 8 2 Le NE 2 ST 4 2ml JO 2 1 EE mWp 18 1112 51 optional NM 5 Outlet alve V6 Valve V4 Sample Pump P 950 Sal 2 RY 0 1 50ml min NO Seeclon 1 0MPa 10 bar 145 psi RN Mi 5 1 Figertight connector 1 16 18 1112 55 4 A PEEK 0 75mmX20cm Figertight connector 1 16 18 1112 55 p 0 0 Figertight connector 1 16 18 1112 55 3 7 7 3 PEEK 0 75mmX50cm Tubing connector for 1 16 o d tubing 18 1127 07
67. Pump SampleFlow 1mMmin Execute Continue 10 1 SampleFlow 10ml min 5ml min Execute 11 3 6 End 12 7 4 4 1 2 5ml Manual Flowpath InjectionValve Inject Execute 3 Manual Pump SampleFlow 2mMmin 4 5ml End Injection Technique Sample Pump Loop Filling 7 5 1MPa 7 5 1 P 950 Direct Load Tubing Kit 18 1152 83
68. Run FE mmwh FwFaEuoh FimcSrpe mr Flrmmilhrmasmh It Em lr mW FF Sm dmmi Seen ried mitas HE Esnsa He HE Scoutin9 Define Show detaillS Flowthrough Start At 1_Start_at EluateFrac StartAt OK 2 Fiwlhenud Fiowthrough_ SME estTube erdTube eriTuhs WeitTube The 1 Run1 Run2 Run3 TargetConcB1 100 100 100 Length of Gradient1 10 15 20 Flowthrough Start At FirstTube NextTube NextTube 1_Start_at FirstTube NextTube NextTube Flowthrough Fraction X1 X2 Eluate Fraction A1 A2 File SaveAs Method Name Techniqdue 001
69. 6 2 1 A11 2 A1 B1 BufferPrep A11 Buffer A2 Acid Base B1 Water B2 Salt UV Cond Pressure A Manual Pump Flow 5ml min Execute B Manual Pump Gradient Target 100 B Execute B2 Manual Flowpath PumpBInlet B2 Execute A2 Manual Pump Gradient Target0 B Execute Manual Flowpath PumpAInlet A2 Execute END 23 J 2 3 Z 2mMmn V1 2 6 3 1
70. 8 1 Frac 950 Frac 950 2m min 2 100ml min 8 1 1 Wma EE TL Keying guide 36 8 1 2
71. HETP 1 5 mg ml 0 1 2 Integrate cm 3 Chromatogram Layout Peak Table Plate hight HETP HETP HETP HETP height equivalent to a theoretical plate HETP L N N 5 54 VR Wh assuming a Gaussian peak N L cm VR Wh VR 0 3M Operations Fraction histogram Cond
72. Next gt Elution Elution Technique Segmented Gradient Number of Segments 3 Wash Out Unbound Sample 2CV Clean After Gradient 5CV ConcB100 Reequilibration after Elution 5CV ConcB 0 VariaDbles Next gt 9 1 Elution Segment 1 3 1 TargetConcentrationB 50 B Length of Gradient 15 CV Fraction Volume 1 ml Update Finish 9 2 Elution Segment 2 3 2 Target ConcentrationB 100 B Length of Gradient 0 CV Fraction Volume 1 ml Next gt 9 3 Elution Segment 3 3 3 TargetConcentrationB 100
73. RHesult OK UNICORN Main Menu Methods 11 5 4 1111 2 RESOURCE S 1 ml Buffer A 50 mM MES pH6 0 B 50 mM MES 1M NaCI pH6 0 4 ml min 5 CV P 950 2 5 5 0 10 20 ml 50ml 0 50 15 CV 100 0 CV 100 3 CV 280 nm 254 nm 214 nm 2 Position 2 Frac 950 18 amp 30mm gt
74. Sample Inlet Pump A Inlet A2 A18 A12 A18 VariableS Show datailsS Wash Out Unbound Sample 5 15m CV 0 1 2 3 146 A18 A2 1M NaOH System Control Manual Flowpath PumpAlnlet A2 Manual Flowpath BufferValve A1 A18 Execute Manual Pump Flow 5mMmin Execute 30 UV END
75. 3 N RI OF A ww EE 2 a 60
76. Customise Report Edit Mode 4 7 LiHHiTms Fmrmh FtrhE EE Ve mmt Lora Pe hrm Fm En ditam Em 5 Properties Setup Chromatogram 6 Selected chromatogram s All Chromatograms OK i i i ll 7 File Save as All Chromatograms Save as dlobal format OK He erm FmFm pi i Fi FF rm mr i brim FPrmW ImFra 10 8 Customise Report Exit 9 Print ChromatogramS Cancel 10 All ChromatogramS 11
77. 9 2 Elution Segment 2 3 2 TargetConcentrationB 100 B Length of Gradient 0 CV Fraction Volume 1 ml Next gt 9 3 Elution Segment 3 3 3 TargetConcentrationB 100 B Length of Gradient 3 CV Fraction Volume 1 ml FiniSh 10 Variables Hie ka IIITL HN LTE fl EE ms i Ir Eg EPR Fm 1 ck ELL IL LETT pd 1 T TFT Fs IH Ti LIL tr bm i 1 E 1 W Wizard VariaDleS Wavelength UV2 UV3 2 3 Value ON OFF 66 lt VariaDleSs gt Wavelength 2 3 Wash Out Unbound Sample Clean after Elution a
78. Frac 900 Frac 900 Outlet Valve OFF Fraction Volume ml Next gt 7 Peak Fractionation Hm mamd om lmrmlmmm mika I Peak Fractionation ON OFF ON Flowthrough Fraction Segmented Gradient FirstTube lt Frac 950 gt Peak IdentificatlOon Level UV Level Slope Variables 9 2 Peak Fraction Volume ml Tube Type 18mm 30mm Start at NextT
79. with ferrule for 1 8 o d tubing 18 1121 18 a Figertight connector 116 18 1112 55 2 PEEK 1 0mmX50cm Figertight connector 1716 18 1112 55 Figertight connector 116 18 1112 55 3 PEEK 0 75mmX15cm Figertight connector 116 18 1112 55 6 Figertight connector 116 18 1112 55 5 Tefzel 0 75mmX130cm Union 5716 female HPLC male 18 1142 08 4 1 6mmX130cm Tubing connector for 18 o 0 tubing 18 1121 17 with ferrule for 1 8 o d tubing 18 1121 18 Union 5716 female HPLC male 18 1142 08 1 8 1 6mmX130cm Tubing connector for 18 o 0 tubing 18 1121 17 with ferrule for 1 8 o 0 tubing 18 1121 18 130cm
80. 5 3 209 1 Pressure limit Manual Alarms amp Mon Alarm_Pressure High Alarm Execute 2 Manual Flowpath ColumnPosition Execute 3 20 Manual Pump Flow 1 2 ExeCcute 1 4 iC 9 4 3 UV Cond Pressure END 21 22 HiPrep AKTA
81. DK 19 9 Rename 1 UNICORN Main Menu MethodSs Results 2 Rename 3 OK 19 10 1 UNICORN Main menu File PrinterSetup 2 142 20 1 AKTAexplorer 10S 100 4
82. 16 UNICORN Main Menu Methods 7 5 11 2 3 6 11 2 RESOURCE S 1 ml Buffer A 50 mM MES pH6 0 B 50 mM MES 1M NaCI pH6 0 4 mMmin 5CV P 950 0 im 0 100 10 CV 15 CV 20 CV 100 0 CV 100 3 CV 280 nm 254 nm 214 nm Position 2 Frac 950 18 amp 30mm gt 1 50 ml 1ml Frac 900
83. Left limit Right limit 6 OK 7 Accept negative Deak OK 7 115 14 6 6 1 Integrate Edit Baseline 2 2 Select Baseline to Edit OK 3 PoOjint list EE EEE Point mode Point list Zoom mode
84. WVVizard VariaDbles Wavelength UV2 UV3 2 3 Value ON OFF Enter lt VariaDleSs gt Wavelength 2 3 PeakFrac Parameters Slope Slope Variables Show detalls Frac 950 gt Variables Frac 950 LEE EL TE EL EEE EE LE LT LE EL ET HE HIITTI T HE HE THEE E INETTH 1 1 1 1 1 1 EE EE ET EL LL LT EEEEEE i T 1 IE 1 IE IT IT Tl EL LL LL EE EE LE LE HE IT TL Rack
85. es mma CreatedCurve cw lee 124 125 14 13 4 Properties Curve Style and Colour Evaluation _ 1 Operations Activity Histogram 3 OK 126 127 lt Frac 900 gt 15 1 2 20 A11 A2 B1 B2 Manual Pump Flow 2mmin Execute Manual Frac Fractionation 900 FracSize 2ml Execute
86. 1 13 3 3 Manual Execute HE FA EE a TT FA 1 2 END End Method Save partial result OK 95 7 Evaluation ee n EL pe 14 1 1 Evaluation 2 File Open 3 OK 14 2 1 2 ProDperties 3 Chromatogram Layout Le Sle red Ck Ei Te Leprat Lia Hesiet Deejimmms Ta ET Le Pa Fm H
87. Elution Segmented gradient HIC System Pump Direct Loadin9 B2 A B 4 100 B 0 9 B 1 Method Editor File New New Method Wizard K 1 2 2 Technique HIC Column RESOURCE Eth 1 ml Column Position Position 2 Next gt 3 Wavelength UV1 280 nm 2 3 Variables 10 Pump A Inlet A1 Pump B Inlet B1 Buffer Valve A Inlet A11 Start Concentration B 0 B Equilibration Volume 5CV CV Next gt
88. Fraction Volume ml Next gt Elution Fractionation Frac 950 gt be nl md hn pmmm mm mm mm _ Heiw a _ Fixed Volume Fractionation Frac 950 Frac 950 Outlet Valve OFF 45 46 3 Fraction Volume ml Tube Type 18mm 30mm Start at NextTube FirstTube 1 Frac 950 Elution Fractionation 18 amp 30mm Tube 18mm Flowthrough Elution Fractionation Start at NextTube 18 amp 30mm Tube 30mm Flowthrough Elution Fractionation Start at FirstTube lt Frac 900 gt I ir TH Fixed Volume Fractionation
89. Resut CN Fn SEAL TE 17 Me HN gnu crore ii ms Bld La mm CL TRAT LEIHNDITIHH ED gr im Cie ei pe Fd Lm ro Lr MN rari Ee LE mem EN ee lr FmrrhmmEnahld SP Ee Em Ls 11 I mT en EE Hm i tt LE Fm hal per F1 Hi rt ee 13 3 13 3 1 SysStem Control 13 3 28 a oF Run Hold Continue Pause Continue Pause Continue Hold Pause End
90. 1 eg TargetConcentrationB 50 B Length of Gradient 20 CV Fraction Volume 1 ml Update Next gt 8 2 Elution Segment 2 3 2 Target ConcentrationB 100 B Length of Gradient 0 CV Fraction Volume 1 ml Next gt O ma Target ConcentrationB 100 B Length of Gradient 3 CV Fraction Volume 1 ml FIinish 9 Variables ia ee a dy 1 la om Tim i FE i i Nm 1 me 1 mm Ne mr ee Wari a EE gsa mm i wm i LL EE Ll ed Fw 1 mm EE EN bi 1 ma LF 1 Li a ms i rp Eri W Nil Fg i Wizard Variables Wavelength UV2 UVS3 2 3 Value
91. X Y 4 X Y 5 Marker 1 CurveS PropertieS 2 Curve Style and Color TE Tl ia Eh a rn WE 1 a ET IL_ Lim1 T Fsamn mm 3 4 OK Y Y 1 CurveS PropertieS 2 Y AxIS HE Na Thee arm Th Cav Tiina Cu es te F fed 3 Fixed
92. pH 1 UNICORN Main Menu 9 pH pH 2 Methods Results pH 3 New Folder Create New Folder 4 DK 19 1 MethodS ReSults 2 Open dd 1 Methods 2 Run 136 13 19 3
93. 150ml 2MPa Execute Manual Flowpath InjectionValve Inject Execute Manual Pump Flow 10mmin Execute End P 950 Inject 8 9 Manual Flowpath InjectionValve Inject Execute Pump SampleFlow 10mlmin 5mMmin Execute 1 2 Pause 969 Pump SampleFlow 50mlmin 25mlmin Execute Continue 2 4 Pause
94. Elution Technique Segmented Gradient Number of Segments 3 Wash Out Unbound Sample 2CV Clean After Gradient 5CV ConcB1009 Reedquilibration after Elution 5CV ConcB 0 VarlableS Next gt 9 1 Elution Segment 1 3 1 TargetConcentrationB 100 B Length of Gradient 10 CV Fraction Volume 1 ml Update Next gt 9 2 Elution Segment2 3 2 TargetConcentrationB 100 B Length of Gradient 0 CV Fraction Volume 1 ml Next gt 9 3 Elution Segment3 3 3 TargetConcentrationB 100 B Length of Gradient 3 CV Fraction Volume 1 ml Next gt 10 Vari
95. Manual Flowpath SampleValve S1 5 Execute 6 Pump 1 SampleFlow 2mMmin Execute 10 0 35ml End 7 File Instant Run Use Template 8 Purgde Sample Pump Run 9 Next gt 10 START 2 7 5 3 P 34 1 2 Manual Flowpath InjectionValve Waste Execute 3 Manual Pump SampleFlow 10mMmin 5mMmin Execute 1 2 Pause 4 969 Manual Pump SampleFlow 50mlmin 25mmin Execute Continue 2 4 Pause 5 6 Manual Pump SampleFlow 1ml min Execute Continue 5 7 Manual Pump SampleFlow 10mlmin 5m min Execute 3
96. System Control Manual Alarms amp Mon Alarm_Pressure High Alarm ExeCcute 10 50ml 4MPa 150ml 2MPa Inject Manual Flowpath InjectionValve Inject Execute Manual Pump Flow 5mMmin Execute 0ml END Union Luer female 1 16 male PEEK 18 1112 51 2 3 5m 9 12 0 1 1 2
97. 4 Fm We ha Fh 1 Eia Irie rrim mrrmrfm TE Larn oh EEEEFI rd EE LE Ff ET ma Ee 1 Injection Techniqdue Sample Pump Loop Filling P 950 Fill Loop with 0 2 ml 85M Empty Loop With 0 2 ml 2 3 Sample Inlet S1 S 8 Rinse Sample Loop 0 5 ml 1 6 Next gt 5 Frac Frac 950 gt Frac 950 Rac 18 amp 30mm Tubes Flowthrough Fractionation Frac 950 Fraction Volume 5 ml Tube Type 30 mm Start at First tube lt Frac 900 gt Flowthrough Fractionation Frac 900 Fraction Volume 5 ml Next gt 6 Elution Fractionation Frac 950 gt Fixed Volume Fractionation Frac 950 Fraction Volume 1 ml Tube Type 18 mm Start at First Tube
98. Frac 900 gt Fixed Volume Fractionation Frac 900 Fraction Volume 1 ml Next gt 7 PeakFractionation Peak Fractionation OFF Next gt 8 Elution Elution Technique Segmented Gradient Number of Segments 2 Wash Out Unbound Sample 2CV Clean after Elution 5CV at 100 B Reeduilibration after Elution 5CV Variables Next gt 9 1 Elution Segment 1 2 1 Target ConcentrationB 100 B Length of Gradient 10 CV Fraction Volume 1 ml Next gt 9 2 Elution Segment 2 2 2 Target Concentration B 100 B Length of Gradient 5 CV Fraction Volume 1 ml UuDQdate FiniSh
99. 100 5 Searchnh 6 7 OUK 1 File Open ChromatogramsS ill ET re mm Tl 2 Resull 3 Available chromatograms 1 1 Select gt gt 4 2 3 5 OK 14 2 8 1 Windows Tile Windows Cascade sm mi PD 101 2 Open
100. 2 20 20 209 pH pH 1 209 2 System Control Manual Pump PumpWash Explorer A11 ON ON ON Execute 3 Manual Pump SystemWash 40ml explorer 10S 75ml explorer 100 ExeCcute 4 S1 209 Manual Pump SampleFlow 2mMmin Execute 5 END 16 2 1
101. Filter Methods Results 1 Filter VieW mode 1 VieW 2 2 a 02 Details _ List 1 lex _ Large ICOnS IGX iex Small lcons jex jex 21GX 4 jex 3 Filter fitered iex 4 Filter Filter Defaultt OK 19 4 1 Find 2 Find fileS Name
102. UNICORN Main menu 6 CloSe 19 5 2HD DOS V 1 44MB 2 1 A 2 Windows My Computer 3 3 1 2 Floppy A 4 Format 5 Start 6 OK 7 Format complete OK 8 CloSe 9 My Computer X 140 19 6 Copy to external 1 UNICORN Main Menu Methods
103. 15 File SaveAs Method Name Technidue 001 RHesult K 16 UNICORN Main Menu Methods 85 7 System Control 12 1 12 1 1 NN DAN 9 LOAD
104. Gml min Accumulator Tube Drop 1 OutletValveF2 Accumulator 2 5mm 3 Knurled screw Finger tight connector Capillary end projection 5mm Capillary projection indication on DropSync unit Gml min Accumulator 1 OutletValveF2 lower DOrt 2 0 75mm 250mm UDD6er port 3 5mm 4 Knurled screw
105. Template 9 1 1 Variables pO File Save As 7 FfFRk SN 10 1 2 2 Superdex 75 HR 10 30 50mM pH7 2 150mM NaCI 0 5 ml min 2 CV 0 2 ml 1 lsocratic 1 5 CV 280 nm 254 nm 214nm Position 2 1ml 12mm Frac 950 1 Method Editor File NewW New Method Wizard OK 2 Technidue Size Exclusion Column Superdex 75 HR 10 30
106. Wizard Fraction order 4 Method Information Method _ Duration 10 mrm ls in tsmrm rl te File SaveAs Method name Technidue 001 Resull OK 55 11 UNICORN Main Menu Methods 10 2 16 RESOURCE S 1ml 4 ml min
107. SN 29 7 5 CC 29 IEEE SCS 29 sss 30 753 Ac A 34 7 6 _P 950 ki 34 8 ui 36 8 1 _Frac 950 36 8 1 1 kk 36 8 1 2 37 8 1 3 kk 38 8 1 4 DropSyncro Accumulator i 39 8 2 Frac 900 kk 40 9 42 9 1 kuku 42 2 Sion 52 UN P RR NC 9 10 1 0 TI I 1ICTIiiiIiy y f 2 53 10 2 56 US 0 0 EE eo 60 11 kk 64 oo 64 11 2 ku 65 2 OH 0 65 11 2 2 71 11 2 3
108. pH 2 3 4 5 6 D7 8 5 0 1 2 AIEX mix AIEX 5 1 methyLpiperazine 9 Bis Tris 6 0 1 Piperazine 6 6 7 Tris NOB 1 methyLPiperazine 8 6 10 3 Piperazine 9 4 10 5 2 14 15 16 17 18 19 HO HH1 12 2 3 4 6 8 9 10 11 12 CIEX mix UE mi lt Phosphate 2 2 3 0 Formate 5 4 5 Citrate 6 62 Acetate 4 0 MES 9 Phosphate 6 4 4 HEPES SM9 0 Bicine 7 0 9 0 2 14 15 16 I 18 19 HO 1 12 64 pH HCI NaOH NaCl 4 pH lr Buffer 1 Salt All Acid Base i A2 Bl 11 2 UNICORN BufferPrep pH
109. 0 B mS cm 4 Monitors SetCondScale100 2 100 B mS cm 5 OK 6 Set Parameter To Default Value 134 18 18 1 FR 902 0 2MPa 1 A 2 2 System Calibrate Monitor P900Press ml 1 TFT Em FI i imsmm Pi mw 3 Start Calibrate 4 18 2 pH pH
110. 12 lt b UNICORN Main Me Melhod Editar untitled a Ewaluation 5nustem Lnhtrnl 1 Gust UNICORN Main Menu Methods 9 2 Method EditOor Wizard 9 1 File File Open OK File Saveas Template Et Block Vsaw Help 1 Hew Ce my Dpar Chl Llpmm ae Melhpd wiggd rt Ll Se Clris Ta ir et ea Epat F Eh Frir Cr Ew Ch wm nl TemmnLsle cm mm Name Technique OK
111. 12 2 0 hethod Editor unhtled 1 System Cortral 1 Smst E yaluahinh Epler per Hename Copu ce Delete Copy to Ewtermal 3 Close 4 5 5 1 Manual Manual 5 2 Flowpath Manual Flowpath 5 3 Sample ValvVe Manual Flowpath SampleValve 5 4 S1 Manual Flowpath SampleValve S1 5 5 Execute Wa TTY ew
112. 4 W 2 Ne i Q a
113. PreVvieWw 14 4 3 1 Save AS 2 Save as dlobal format Save as default report format ae Immat Ferm Hm HE LE pi tr op ii mi mr ol bom 5 HFPrmW ImFrg 3 OK 4 Exit 5 ClOoSe 109 14 5 Resut View Documentation NN PrInt UK VariaDles mmm Lejos Emel heer Vi some fmmmmt Hm Hmmmmr Suna Camrr hmvm EmmamrmPapskumr Scouting
114. W1 W2 WS F3 10S PEEK 100 4 LL F3 F3 20 Pump Pump
115. Wizard Variables Reequilibratiton 5 CV FIniSh VariaDleS 9 Variables rl rm Ea i i rm ie re Hkaurk LL a a mr ma ee EE ei re Ei a LE Fm Pp L lmms ed li Fx a a i mm 1 a ie i ra ir Fm mr a mu i Em 1 i Fm mmmdPen ima Fu 7 Eh br Lire ll i msi ee EE Firm jmM EF Wizard VariaDbles 48 Value VariableS gt Wavelength 2 3 Wash Out Unbound Sample Clean after Elution
116. 5 Create New User OK User Setup 2 User Edit 6 User properties Access Folders 7 User properties Systems System1 13 i re mlm mmmihn mm bg 8 OK User SetuDp ClOoSe Method Results Logon 9 12 Logon 9 Logon Logoff Yes 10 Leave Control of System1 Lene Cr of rinm 1 Li
117. Pause Continue rm El I na se 1 na 1 1 mi LL 1 a J i Pd Ta ECimT a Ti itis 3 Te LT 1 Ta Sie 40 8 5 5 mm Lock Knob et 5 mm ee a gt Sa lt lt gt PEEK 5 mm
118. Wash Out Unbound Sample Clean after Elution at 100 2 B Reequilibration PeakFrac Parameters Slope Slope VariableSs Show details 83 64 Frac 950 gt VariaDbleS Frac 950 Rack Wizard 18 amp 30mm Tubes Fraction order 4 VariableS Scoutin9 Fill loop with Empty Loop with OK Scouting Add Insert Run1 Run2 Run3 Run4 Fill loop with 40
119. 1 No 1 Sample volume and type 2 Default text 3 Replace 4 Start protocol gt Method Queue Start protocol Variables Result Scouting Resut Text Method QuestionSs
120. 3 UV Cond Pressure END pH pH KCI 129 15 7 130 UNICORN Main Menu File Quit program YES Leave Control OK Windows Start Shut Down HH tart hd mn Wm Ra HB TR LE SU af Im et dow th cempum Heaskmt the compiule Ciuse ol prame and Inn on ssa de usem E FF 76 16 1
121. 73 74 10 Variables Wizard Variables Wavelength UV2 UV3 2 3 Value ON OFF Enter lt VariaDles gt Wavelengdth 2 3 Wash Out Unbound Sample Clean after Elution Reequilibratiton PeakFrac Parameters Slope Slope VariableS Show detalls Frac 950 gt Variables Frac 950 Rack
122. ML S8 1 1 S8 S1 S7 Manual Flowpath SampleValve S1 Execute Pump SampleFlow 2mlMmin Execute 10 0 35ml S1 End 3 4 S2 S7 Flowpath SampleValve S8 Execute Pump 1 SampleFlow 2ml min Execute 1 nd Flowpath InjectionValve Inject Execute Pump Flow 2ml mn Execute 5 End Injection Technidue Sample Pump Loop Filling
123. S6 wash Vol 5 Variables 1 15 SampleFlow 0 00 ml min 2 15 SampleFlow 0 00 ml min 7 2 5 6 a Em Brt Han 0 00 SampleFlow 50 ml min lag si ll 0 00 SampleFlow 25 ml min mpgn mm ml fim mmEagi me HH ala ia tr 2 Wash Time SampleFlow 15 BT alan mm 0 00 ImMmin 1 mm le i nl ni Bammrieralmm 7 Blsrk Wa Wh_ Etharsl 4 Wash Time SampleFlow 0 00 ImMminl mm mmm ai SamalsFinw 25 EIHS_Flsm lmirmml aaah Tire misFinm 0 80 imimmi 7 65 SampleFlow 10 ml min 7 65 SampleFlow 5 mil min 10 65 End method 13 65 End method Variables variable name SampleFlow1 SampleFlow2 8 Variables Run Setup P 32 5 7 Variable Name
124. 1 50 ml 1ml Frac 900 1 Method Editor File New New Method Wizard OK 2 Technidue Cation Exchange Column RESOURCE S 1 ml Column Position Position 2 BufferPrep OFF Next gt 3 Wavelength UV1 1 280 nm 2 3 Variables Pump A Inlet A1 Pump B Inlet B1 Buffer Valve A Inlet A11 Start Concentration B 0 B Equilibration Volume 5CV CV A 3 5CV Next gt 6 1 82 4 Injection technique Sample Pump Loop Filling P 950 Fill Loop with 40 ml Empty Loop With 2 5 ml Sample Inlet S1
125. J C_ 4 AN AKTAdesign UO ke 9S 1 1 AKTAexplorer kk 5 oo 10 2 1 UNICORN eee 10 2 2 5 5 11 2 2 1 11 2 2 2 kk 12 EE 13 5 16 3 1 ek 16 EE EE 16 3 3 kk 16 3 4 un 17 4 esses 19 41 kk 19 42 DST RIIIIY8SN 55IIAIIEIII 19 4 3 es 19 5 ee 0 3 2 Mi 20 FPLC 20 AKTA 20 a 2 20 PI PPP 20 EE Ek E E ss 21 5 37 A 21 6 kk 3 AE EE 23 6 2 kk 23 7 es 4 24 SAMDIEPR MD PU UR RE 24 7 9 a 25 73 1 i 25 7 3 2 ka 26 7 4 ki 27 7 27 7 4 2 es 28 se 28 744
126. Frac 950 Fraction Volume 1 ml Tube Type 18 mm Start at First Tube Frac 900 gt Fixed Volume Fractionation Frac 900 Fraction Volume 1 ml Next gt Peak Fractionation Peak Fractionation OFF Next gt 6 8 Elution Elution Technique Segmented Gradient Number of Segments 3 Wash Out Unbound Sample 2CV Clean After Gradient 5CV ConcB100 Reequilibration after Elution 5CV ConcB 0 pH Variables Next gt 9 1 Elution Segment1 3 1 Target ConcentrationB 50 B Length of Gradient 15 CV Fraction Volume 1 ml Update Next gt
127. 2 Fm udFlg ia 1 IT NetTube HedTube Tubetiumbesid 1 Run1 7 0 Flowthrough Start at FirstTube 1 Start at FirstTube Flowthrough Fraction X1 X2 Run2 Run3 6 0 5 0 NextTube NextTube NextTube NextTube NextTube NextTube Run4 4 0 Eluate Fraction A1 A2 HE ai Pe kha Tam Ff PL LE HL HE i I Ee Ec _ eiF wal EL i I PT Cun Hi File SaveAs Method name Technidue 001 Resultl K 16 UNICORN Main Menu Methods 11 2 2
128. pH Cell pH 0 25mm 105 ml 0 5mm 301 uil 423 pl 87 AKTAexplorer 100 12 4 Accumulator pH Cell pH 1 0 5mm 448 pl 570 Hl System Control 0 75mm 775 Hl 924 pl File Run 1 0mm 1232 ul 1418 pl _ Accumulator pH Cell pH 0 5mm 297 ml 419 ul 0 75mm 624 pl 773 Hl 1 0mm 1081 ul 1267 ml lt Frac 900 gt 1 2 5mm 3 Outlet Valve F3 2 PEEK J OK 9 4
129. 1 Method Editor File New 2 New Method Use 1 Template Template j Purge Sample PumDp ET FI TE FET TF Fl a I Fh mml ed a Es ee a i 3 OK 5 i ss ms a 4 File Save ASs gt Purge Sample Pump Step1 S6 10mMmin 1 5mMmin 2 Step2 S7 50mMmin 6 25ml min 4 Step3 S6 10mMmin 3 5mMmin 6 1 3 4 Text Mode
130. 97 X i 98 14 2 3 sk 98 14 2 4 lt 99 14 2 5 lt 99 E E EkE 100 14 2 7 es 100 100 2 0 ON SETETETKT TK KKKTKK 5 5 101 14 2 8 101 14299 SpA 102 kk 102 OOo 104 105 14 2 10 kk 105 2 Fibs esaaaaaiindeiabsaeaaia And ei a nae ea 106 kk 106 lt 106 14
131. Variables mm a 1 EN Il IIT SEE rs ata Ha Ei i mam i mmmm Fammmm i i Td LA Hr IS sitm IF HILL meE 9 File Save As 7 5 3 P 31 Sample Pump Direct Loading 7 6 P 950 P 950 Purge Kit P 950 P
132. uF RTE rl SE lame iFP mm 4 Logon InformattlOn Password OK Enter LE a PEE el Fad al 1 li 5 Windows FE i 6 UNICORN a UNIEDEN 7 Logon OD User name default default Car OK 2 3 13 8 System Information Ilnstruments are ready Clear All i 1 9 UNICORN Main Menu lt UNICORN Main Me Method Editor hllgd 1 System Control 1 mst Ewa
133. 8 1 4 DropSyncro Accumulator DropSyncro Accumulator Tube Tube DropSyncro Accumulator OFF 2ml min 2 2mlmin IM System Control System Settings Specials Tube Change 39 8 2 Frac 900 fr JNI FE 1 1 a Ln Ld i LT
134. P 950 Frac 950 p12 B S8 Rinse Sample Loop 0 5 Next gt 5 Frac 950 lt Frac 950 gt Frac 950 Rack 18 amp 30mm Tubes Flowthrough Fractionation Frac 950 Fraction Volume 50 ml Flrouthrough Fraction Volume Tube Type 30 mm Start at First tube Frac 900 gt Flowthrough Fractionation Frac 900 Fraction Volume 5 ml Next gt Elution Fractionation Frac 950 gt Fixed Volume Fractionation Frac 950 Fraction Volume 1 ml Tube Type 18 mm Start at First tube Frac 900 gt Fixed Volume Fractionation Frac 900 Fraction Volume 1 ml Next gt Peak Fractionation Peak Fractionation OFF
135. gt Variables Frac 950 Rack Wizard 18 amp 30mm Tubes Fraction Order Serpentine row 4 File SaveAs Method Name Technidue 001 Resultl OK UNICORN Main Menu Methods 63 17 11 1 BufferPrep pH pH UNICORN De
136. 12ml explorer 100 V1 3 Load Inject Waste Pumpwash 0 Column 5 Column Pump Sample 6 syringe Waste Waste Load V1 2 6 explorer 10S 10ul 100ul 1ml explorer 100 500ul 2ml
137. 5 amp 1 Edit Text Mode 2 3 Edit Text Mode 14 3 106 1 File Print Print Chromatograms EEI hh kate momatnawhmw ff am Hm ra Pe Li LT cma tm 2 Print for
138. 6 End 7 5 3 P 34 x x S1 S8 0 75mm 10ml min P 950 Purge Sample PummDp 2 S6 S7 969 31
139. V3 AN Kb AKTAexplorer 10S 19 5 5 1 1716 male PEEK FPLC a M6 Union M6 female 1 16 male PEEK 18 1112 58 8 SRTC 2 19 2143 01 AKTA SRTC 2 1 16 0 5mm PEEK 100 0 75mm PEEK Fingertight Connector 1 16 18 1112 55 10 HiTrap M6 M6 0 5mm PEEK 100 0 75mm PEEK Fingertight Connector 1716 18 1112 55 10 Union 1 16 female M6 male PEEK 18 1112 57 6
140. 3 Chromatogram Layout 4 Curve Style and Color 5 Retention Number Peak Name No Peak Label 1 2 ProDperties 3 Chromatogram Layout 4 Peak TableS 5 Filter PeakS 6 OK Peak Integrate Integrate PeakIntegrate 2 Reject DeakS Peak Integrate TT LE FE NE ja mm ha ET TTF a Lm 3 Peak WinQOwW 4 Peak window 5
141. 3 Instruction box Breakpoint Instructions Parameters Change Replace Change BreakDoint hr Le Hi lm mu td am IT Nm EE Fm rmrirr Fr 1 gE nl TE aa uma TE Es i a HI Har HO MEET Hn DN hmm ERD RT BE UE Em bet el red Eicri Replace Breakpoint eplsce Fi Le i LE He er Fd LE Hr Ce Td er HD Fr ui co Sin Il rw 1 TT mmml Io Huimrm 1 hi mai SN Frc tke a ee EA OF rT Hr FT re Ed ss 5 Lr srmm 0 3 1 3 1 Text 2 3 Instruction box Delete 149 GE Healthcare _
142. T EE 5 5 ee 143 20 1 1 kk 143 20 1 1 1 144 20 1 1 2 kk 145 20 1 2 i 145 20121 i 145 20 1 2 2 kk 146 20 1 2 3 kk 146 20 2 kk 147 20 3 148 20 3 1 Text Instruction eee 148 20 3 1 1 kk 149 20312 SAD A A 149 20 3 1 3 kk 149 7 AKTAexplorer 1 1 AKTAexplorer UNICORN
143. aa og 5 Start 6O 89 7 9 mm ss i a hn ET LE HU LT HiFFH EE 13 1 1 View JJ Panes 2 Run data CurVeS OK 3 amp 13 5 13 2 1 Run data 1 Run data 90 2 Properties Run Data Groups TT Cust Tp os Hi eis Ree nn fi Enkin
144. ee TE ha Pa ri Fie mm hm 3 Logon 1 UNICORN Main Menu 2 Administration UserSetup RIO He Meer EL aeW Ta La 5 pte Hr Maem Fi Li En ir tia ld 3 User Setup New Create New User Hae tion ll re en EL hm End Li ha User Name a ra tst Full name sm Position Me li mm 4 Home folder New Create New Folder Folder name OK
145. 14 6 7 1 Chromatogram Layout Curve Style and ColOur NumDper Retention 2 Integrate Edit Peak Table Select Peak table to Edit 3 OK 4 OK EE i i 1 11 Delete peak
146. 2 1 File Report 2 Generate ReDort TE TI ul end Lire Te ET Hur ur ed hl We T mf He Era nti fhe ee tn 1k Ti EL 3 4 Generate ReDort Edit 108 5 Add Page Delete Page LIHihTma Fmhmhi Frh Fr a Vi rt i Em e In 6 Insert 7 Properties Setup Chromatogram 8 Settings Layout Current Define Y AxIS Y 4 9 FontS OK
147. 5 CV 1 ml 0 50 B 20 CV 50 100 B 0 CV 100 B 3 CV 280 nm 254 nm 214nm Position 2 Frac 950 18 amp 30mm gt 1 50ml 1 ml Frac 900 Method Editor File New New Method Wizard OK Techniqdue Cation Exchange Column RESOURCE S 1 ml Column Position Position 2 BufferPrep OFF Next gt Wavelength UV1 1 280 nm 2 3 Variables 9 Pump A Inlet A1 Pump B Inlet B1 Buffer Valve A Inlet A11 Start Concentration B 0 B A Equilibration Volume 5 CV CV A
148. A11 A2 Manual Flowpath PumpBInlet B2 Execute B1 B2 7 2 4 A2 B2 3 4 1 System Control Run data CurveS OK CT Fe mri Lact ee is 2 A A11 Manual Pump Flow 5ml min Execute 3 Pressure 4 B B1 Manual Pump Gradient Target 100 B Execute 5 Pressure 6 B B2 Manual Flowpath PumpBInlet B2 ExeCcute 7
149. EKE k k 5 5 131 16 3 131 16 4 131 16 5 132 16 6 i 132 kk 133 17 1 133 72 DSTD GA 134 17 3 Conmd a 134 Lucccccciciiiitiittittttttanntaaneannannnencenne neers 135 18 1 135 1 pH 135 Main menu kk 137 191 Na i 137 19 2 kk 137 i 137 AAcjAckcCG 137 19 3 kk 138 oi 138 oe 138 Elio 139 194 DD 139 19 5 140 0 Copy to exletnab ee 141 19 7 Copy from external 141 19 8 kk 142 19 9 Rename 142 19 10 kk 142 oo 149 0 0
150. ResultS 2 CODpy to external 3 Copy to 3 1 2 Floppy A Save Copy to external Result ZID zip Copy from external 19 7 Copy from external 1 UNICORN Main Menu Methods Results 2 3 CODy from external 3 Copy from 3 1 2 Floppy A 4 K
151. Science Program Web Www gelifesciences co jp e mail Web 2011 GE GE A uka 169 0073 3 25 1 ISO 9001 2008 TEL 03 5331 9336 _ FAX 03 5331 9370 e Imall Tech JP ge com
152. Wizard 18 amp 30mm Tubes Fraction order Serpentine row 4 VariaDbleS Scoutin9 Column Column position OK Scouting Add Insert Run1 Run2 Run3 Column Resource ETH Resource ISO Resource PHE Column position Position2 Position3 Position4 Column Value uDdate YeS lt 13 14 Fraction Frac 950 gt Flouthrough Fraction Eluate Fraction Run LT n Fir Fip TTT L
153. with ferrule for 1 16 o d tubing 18 1127 06 3 W2 Figertight connector 1 16 18 1112 55 5 EB NN 6 Tefzel 0 75mmX130cm 7 4 Union 5 16 female HPLC male 18 1142 08 1 6mmX130cm Tubing connector for 1 8 o d tubing 18 1121 17 C 1MPa with ferrule for 1 8 o d tubing 18 1121 18 75 5 Figertight connector 1 16 18 1112 55 1 8 1 Tefzel 0 75mmX320cm Pump P 950 Outlet connection Tubing connector for 1716 o d tubing with 2 ferrule for 1 16 o d tubing _ Union 5 16 female HPLC male TR Tubing connector for 1 8 o d tubing with rE a ferrule for 1 8 o d tubing Inlet connection 24 2 7 3 6 System Control 26 7 4 7 4 1 A
154. 0 1ml 0 50 B 15CV 50 100 B 0CV 100 B 3 CV 280 nm 254 nm 214nm Position 2 Frac 950 18 amp 30mm gt 1 50ml 1ml Frac 900 Method Editor File New New Method Wizard OK TTT mm F om PE EE i m om mm Techniqdue Cation Exchange Column RESOURCE S 1 ml Column Position Position 2 BufferPrep ON BufferPrep Recipe 3 0 7 5 pH CIEX BufferPrep pH 7 Next gt 3 Wavelength UV1 1 280 nm 2 3 Variables 10 Buffer Valve A Inlet A11 Start Concentration B 0 B Equilibration Volume 5CV CV A
155. 3 4 New GrOUuD 5 OK 13 2 2 Curves EL nn np a mn a mn a mn En 1 Curves 2 Properties Curves rE LT ms Ceti Fi ar LFei LT er i Tas i PIT Ti Firm rE rT TE a ie Te me Fh ET TI LT 1TT ha 3 Select All 4 OK 91 92 1 CurveS Marker 2 3
156. Chromatogram Layout Header 2 Questions 3 ET EE 4 5 OK 14 2 5 1 Chromatogram Layout Layout LiDrary 2 Save Current Layout aS 3 OK Save as default 4 OK 99 14 2 6 1 2 3
157. br rE mm HT EL Hm Li Ti mr Fc lt Frac 950 gt VariaDleSs Frac 950 Rack Wizard Fraction order 4 VariaDleS Scoutin9 Show details Target Conc B1 Length of Gradient1 OK Add Insert Run1 Run2 Run3 TargetConcB1 100 100 100 Length of Gradient1 10 15 20 9 60 lt 13 14 Fraction Frac 950 gt Flowthrough Fraction Eluate Fraction
158. 2 1 4 5 END 15 4 8 Frac 950 gt 1 Manual Pump Flow 2ml min Execute 2 Manual Flowpath OutletValve FracF2 Execute 15 6 pH 3 Accumulator Accumulator Manual Frac AccumulatorWash Strokes 5 Execute 4 Accumulator Manual Flowpath OutletValve F3 Execute 5 3 END 128 Pressure limit Manual Alarms amp Mon Alarm_Pressure High Alarm ExeCcute Manual Flowpath ColumnPosition Execute _ Manual Pump Flow Execute
159. 5 Resut Results 6 Method Method System Connection 141 hd pa hee Crc hen mr1 re a rs Cc ri mm mr coe be 7 Method file Systems system 1 8 OK Copy from external zip CODy zip 19 8 1 UNICORN Main Menu Methods Results 2 Ctr 2 Delete 3
160. HPLC male 18 1142 08 1 6mmX130cm Tubing connector for 18 o 0 tubing 18 1121 17 with ferrule for 1 8 o0 9 i 18 1121 18 Tubing connector for 1 8 o d tubing 18 1121 17 with ferrule for 1 8 o d tubing 18 1121 18 Pump P 950 Outlet connection Fingertight connector 1 16 js 1 Union 5 16 female HPLC male Tubing connector for 1 8 o d tubing with 3 a ferrule for 1 8 o d tubing Inlet connection 27 7 4 8 1 7 4 3 28 0 2 6 Manual Alarms amp Mon Alarm_Pressure High Alarm 4MPa
161. Result IT Fm m Wr Sci Rat Kear Sam hen Bars Maieree Harvei Fra Fhe 110 NoteS Notes Evaluation NoteS CaliDratlOn Ei r 1 i i yi im Ges mi nim i i en mmf 111 Logbook 14 6 Integrate 14 6 1 Integrate HE BE Ma rene tire 1 Integrate PeakIntegrate la EE i EF 1 et cot te 2 EvaluatIOn LODE Evaluation Drocedures ma 1 EE mi EE i 3 Target peak table A H
162. at 100 2 B Reequilibration PeakFrac Parameters Slope Slope Variables Show detaills 10 Run Set up lt Frac 950 gt Frac 950 0 HI ams mm me a Fre Nm a er AT ex Rack Wizard Fraction order 4 lt Gradient gt X lt Questions gt H HM Nh ll 49 50
163. 0 0 0 Empty Loop with 2 5 5 10 20 13 14 Fraction Frac 950 gt FlOowthrough Fraction Eluate Fraction Run ee a i FE mr Fheihreh ml Eip cnn ewh h Ea FF 1 Tukm_Tmpm 1 Hmm om wm 1 moeml rr mh ol red 1 Fr hc ela HT UE He Scoutin9 Define Show detallsS Flowthrough Start At 1_Start_at EluateFrac StartAt K MELm MamlLirme Tbg umbmf 11 Run1 Run2 Run3 Run4 Fill loop with 40 0 0 0 Empty Loop with 2 5 5 10 20 Flowthrough Start At FirstTube NextTube NextTube NextTube 1 Start at FirstTube NextTube NextTube NextTube Flowthrough Fraction X1 X2 EIlute Fraction A1 A2
164. 3 106 1 107 14 4 es 108 14 4 1 108 14 4 2 es 108 14 4 3 oe 109 14 5 Resultl 110 14 6 Intearate ki 113 14 6 1 Integrate kk 113 14 6 2 es 114 14 6 3 EEII777YY7YY7EYKY777 WW 114 6 Oo 115 14 6 5 FeGKINIeINSIG KT KK KK Eo 115 14 6 6 116 kk 116 kk 117 kk 147 14 6 7 ki 11
165. 4 131 16 5 7 lt Frac 900 gt 19 6067 02 17 1 1 System Control 2 System Settings 18 1154 17 3 Specials FracParameters DelayVol 0 39ml 1 5
166. 4 Baseline Calculate Baseline 5 OK HivTImT Te a 1 IT 113 14 6 2 1 2 ProDperties 3 Chromatogram LayoOut 4 Peak Table TFT Fr EE Tr re i Er i Ti Lr 1 i PE ri 1 Ph lr br 1 re Fl 1 mr EE rH i mE em me Ee i ef ee Fs ei el he 1 ed ET im _ lm Fd i be TR um ig LM 5 Select peak table columns 6 OK 14 6 3 114 1 2 ProDperties
167. 4 OK 2 3 Frac 9850 gt AKTAexplorer 10S Accumulator pH Cell pH 0 25mm 256 ul 0 5mm 451 ul 573 ml Accumulator pH Cell pH 0 25mm 105 uil 0 5mm 301 ul 423 ul AKTAexplorer 100 Accumulator pH Cell pH 0 5mm 448 pl 570 ml 0 75mm 775 Hl 924 pl 1 0mm 1232 ul 1418 pl Accumulator pH Cell pH 0 5mm 297 ul 419 pl 0 75mm 624 nil 773 ml 1 0mm 1081 ul 1267 ul lt Frac 900 gt AKTAexplorer 10S pH Cell pH 0 25mm 105 ml 0 5mm 301 ul 423 u
168. 7 118 lt 118 OCG GO SCRA 118 kk 118 2 CC 119 14 6 8 HETP kk 119 14 7 0 IIIRITYTITIYITOTOTmII 0 IIIOOITIOO 1 119 14 8 a 120 i RN 120 OOOO 55 121 14 90 SID NN 121 14 10 Export kk 122 14 11 i 123 14 12 kk 123 139 CCSDTDSTDODTDSTDS ODADASRCSCSCSCSCSCSC SS 126 ii 128 15 1 kik 128 SO 128 15 3 kk 128 16 17 18 18 20 15 4 kk 128 15 5 kk 129 15 6 _ pH i 129 esse 130 EEE 19 1 S141 131 OO ETE EE
169. 7 3 25 7 4 27 5 29 P 1 2 2mlmin Tube DropSyncro 2ml min Accumulator SystemControl Systems Settings Specials Frac Parameters TubeChange 3 Outlet Valve F2 2 P 37 Accumulator Accumulator 4 System Control Systems Settings Specials Frac Parameters DelayVol AKTAexplorer 10S Accumulator pH Cell pH 0 25mm 256 ul 0 5mm 451 ul 573 ml Accumulator
170. 950 Injection technidue P 950 1 Injection Valve Position Manual Flowpath InjectionValve InjectionValve Position Execute P 950 Injection Valve A Load Bj Inject Ci Waste 34 2 P 950 P 950 Purging port with stop piug Purge tubing connector 3 4 5 5ml 96 6
171. B Length of Gradient 3 CV Fraction Volume 1 ml FIiniSh 10 Variables Fn ee u ELLE II 1 IL EE rs FP Lm PE Hn Per pe a FT pn LEI LE Hm eign a 1 1 1 mm li Fa er a ne i i al a IL EE JE mm L ue i i 1 ma cg a CN EE i LE 1 lIR F F be i mm mp m bn Lie PF ET i pl a Fim as Fmmc a NE mm LE m mn ea IFmmmH I Bn Fa mm eae mml me Fi FE Nr ee EE Wizard VariableSs Wavelength UV2 UV3 2 3 Value ON OFF Enter lt Variables gt Wavelength 2 3
172. Column Position Position 2 Next gt 3 Wavelength UV1 1 280 nm 2 3 Variables 9 Pump A Inlet A1 Pump B Inlet B1 Buffer Valve A Inlet A11 Start Concentration B 0 B A Equilibration Volume 2 2CV Next gt 4 Injection Technidue Manual Empty Loop with 0 3 ml Next gt Frac 950 gt 5 Frac 950 Frac 950 Rack 12mm Tubes Next gt S 6 Elute Fractionation Fixed Volume Fractionation Frac 950 gt Frac 950 Frac 950 Fraction Vo
173. Fractionation OFF Next gt 8 Elution Elution Technique Segmented Gradient Number of Segments 3 Wash Out Unbound Sample 2CV Clean After Gradient 5CV ConcB1009 Reequilibration after Elution 5CV ConcB 0 VariaDles Next gt 8 1 Elution Segment 1 3 1 TargetConcentrationB 50 Length of Gradient 5 Fraction Volume 1 ml Update Next gt 8 2 Elution Segment 2 3 2 TargetConcentrationB 100 Length of Gradient 0 Fraction Volume 1 ml Next gt 8 3 Elution Segment 3 3 3 TargetConcentrationB 100 Length
174. I HE le Tm mg be T 1 IT Hf e 1_Ur3 em ll le 1 mea ml Eo Ei bg 1 Ci 1 1 IH ala Tarn wT te Fa tms 1 96 nn bh a I Cur 14 2 1 1 Chromatogram Layout CurVvVeS 2 3 OK 14 2 2 Y 1 Chromatogram Layout Y AxiS La BE mLOkeu EmW Te Lepr hm Heyiet Ce Mres eat tat Ere a 1 LF Et Ba 0 L bm F Fm im apphy ke ll rho _ mk Lares Hm 2 3 AutO Fixed 4 3 UV All with thiS unit 5 OK 9 X 1 Chromatogram L
175. L lh Fp mh i i cL i 1 _ ubs Tm mm MW rm a Hd 1 hr ha OR UFJ mhla m Ca Ha Scouting9 Define Show detaillS Flowthrough Start At 1_Start_at OK 2 Figmlhanud Finwlhinunh_ TRPG HestT ube MautTube Run1 Run2 Run3 Column RESOURCE ETH RESOURCE ISO RESOURCE PHE Column position Position2 Position3 Position4 FrowthroughStartAt FirstTube NextTube NextTube 1_Start_At FirstTube NextTube NextTube Flowthrough Fraction X1 X2 Eluate Fraction A1 A2 File SaveAs Method name Technique 001 Resultl OK
176. Tm rw am Im Pam mm Fa 1 TT Unlocked OK 11 UNICORN Main Menu did Logon Logoff 14 12 LO9On howm EDO rm LL EET re ba na ar Hm default OK 13 Change PassSword 1 mmmrmms i ms ums mmm ie tr Logon password Old default NeW Confirm SIgnature DaSSWOrd Old default New Logon password Confirm Logon password OK 15 3 System Control System Control
177. ables ms NL EL IL Sn II IL EE rm ms 1 I 1 mi Bm 1 NE ma PT pe ET i Pl dF 1 ms Ch Fu Ne ed ee fa I Te Wizard Variables Wavelength UV2 UV3 2 3 Value ON OFF Enter Variables gt Wavelength 2 3 Wash Out Unbound Sample Clean after Elution at 100 B Reequilibration PeakFrac Parameters Slope Slope VariableS Show DetailsS ET 1 1 mn 1 ll ami 1 hi rn Ly
178. ayout X AxIiS 1 2 X Auto Fixed 3 Adjust retention zero to injection number 0min ml 4 OK 14 2 3 1 Chromatogram Layout Curve Style and ColOur Header Pseftgmmh ron am mm FosmTabke Cue am red Fr D mal 1 Vs mm O tat 1 UV3 Thm DH te 1 Com lesr1 1 ot 1 1 Le MI 1 I ts 1 Prmime HI IT re Helerimm PTE J FF FF Hg i Habe Fa mW Hvac Fm Ff Ce Fy Cee Ne Poek Lahnl Ein Hmrh Amy ko Ml tho re ce Hm 98 2 3 4 OK 14 5 4 1
179. d LE ee re ee EE 00 mm re 1 Sort 2 Name System Methods Size Type Modified Created 138 139 3 TVDe Methods All Folders Method files MethodQueue files Results All Folders Result files Scouting files Date OK 4 Searcn Found folders and fileS ES 5 5
180. em Calibrate Result name changeable Result Result BrOwSse SCOuting start protocol lt BufferPrep gt BufferPrep Wizard lt Method Informatlion gt Method Duratlon File Save As Method name Technidue OK 001 Resultl
181. em 1 me m Fr ee Fe Hmsm varsbhks Fael FmmE Ranl Fmd ai Eo der i LL wa em at Dm mm _ mi Ht Run1 Run2 Run3 Run4 7 0 6 0 5 0 4 0 Scouting Add Insert 69 70 13 14 Frac 950 gt 13 im wm Flawihiegh 1 Eie lcrn mh Fr hc mm Si iain mi as IE 9 Csml Eeh Flowthrough Fraction Eluate Fraction Run Scoutin9 Define Show detailS Flowthrough Start At 1_Start_at EluateFrac StartAt OK BufferPrep pH
182. itkar LihH hmt En mrHurm Emma re Ht a maa Fr RWHrarsmi nHF WW Pe FE Lt Cat Larm Lfeg Lu 123 2 mm CN 5 Draw spline Spline Though UT GF T LT Fl TE FT NE em ara 13 Fm 1 IPV3 2ETrmm HFT 1 Parg aprri rs TE AIT 1 1 rr 5 GF TDD 1_ Ca HH GF TO Peare iT TI FFT Tammm Ma 11 FTTE1LFIgE TH 1 FT 1 ggi LL 3 FTET1 1_5amlrFai 1 FEIHTT1 re kma mr um EF TTT Loh cm mee Lirain Ci Saict hm Ervelit wD FT LT rm UF 1 TB rr FE EA Le ra MEF 1_Lrrmd LE DE FE 1 IITIELE TH IL FTO 1 mm LF rw 11 FT1 Fa lfami 1 rd 14 FT Tu a EF FN Lehrerk ww 1 tw 6 Draw straight to next point 7 OK 8 OK art Lurre aw CUTWE I 4
183. ixed Volume Fractionation Frac 950 Fraction Volume 1 ml Tube Type 18 mm Start at First Tube Frac 900 gt Fixed Volume Fractionation Frac 900 Fraction Volume 1 ml Next gt Peak Fractionation Peak Fractionation OFF Next gt Elution hr g Dm EMF mn En nh a mmr mer FF th HE i rho mmmm em Im Ein 1 Feehlim ET Emi5 Fl 9 Elution Technique Segmented Gradient Number of Segments 3 Wash Out Unbound Sample 2CV Clean After Gradient 5CV ConcB100 Reequilibration after Elution 5CV ConcB 0 VariaDleSs Next gt 8 1 Elution Segment 1 3
184. l AKTAexplorer 100 pH Cell pH 0 5mm 297 ul 419 ml 0 75mm 624 ul 773 ml 1 0mm 1081 ml 1267 ul 132 133 17 10S3 25MPa Explorer 100 10MPa 1 System Control 2 System Settings a 1 TE NE re Ey 3 Alarms AlarmPressure HighAlarm 4MPa Superloop 50 ml 4 OK 17 3 Cond Cond9 4 B 0 1009 1 System Control 2 System Settings hs i TE bE od Fi hie le i a i Pa Ta Ci Ee ut EE 3 Monitors SetCondScale0
185. linefilter Va V7 Valve door Sample Pump P 950 UV cell only in AKTAexplorer 100 Switch Valve Conductivity cell and AKTAexplorer 10S H5 H12 _ H9 H8 V2 pH COND UV G3 G4 Ww G1 G2 ggK A B 0 A3 BS3 A1 A2 B1 gt B2 AN AN lS KB2 W2 AKTAexplorer 10S Aa B1 J B2 V3 G3 G4 G6 Column 2 BS alve G12 G11 G1 G2 NO0 6 V2 Pump Pump L dV A B I Wo pH COND UV es A3 A2 B1 B2 G15 gt gt Sample w vV Vp Pump TE a Fraction G17 Collector Buffer SBIa G14 Outlet Valve V6 Vale V5 Valve V4 MI MI LA RR 82 S462 JJKJK JA UM MM A11 A12A13 A14 A15 A16 A17 A18 S1 S2 S3 S4 85 S6 S7 88 Valve V3 W3 F3 F4F5 F6F7F8 AKTAexplorer 100 A1 8 Al1 A18 M 925 explorer 10S 0 6ml explorer 100 2ml 2ml 5ml
186. luation 10 2 2 1 UNICORN 4 UNICORN Main Menu Method Editor System Control Evaluation gt UNICORN Main Me S Eyaluainn ustem Lnhirn 1 Sust 9 kMlglhnd Editor unhtled UNICORN Main Menu CbUNICORN Main Me Copy Rename Delete Move Melhad Editor untitled Method Editor 1 System Contral 1 Syst System Control Ewaluahiam Evaluation Instant Run UNICORN UNICORN Logon Logoff UNICORN Logon Logoff Method Quene Method Quene Editor Display running Method Quene 11
187. lume 1 ml Tube Type 12mm Start at First Tube Frac 900 gt Frac 900 Frac 900 Fraction Volume 1 ml Outlet Valve Off Next gt 7 Peak Fractionation Peak Fractionation OFF Next gt Eirmi 1 FE LC EE a i TT a leer mT lm WF FT FET MT al LN lm Peak Fractionation ON 9 1 7 8 Elution ELMmei EhmmTmrhrw Em Fi th im md Ft FE TF Er TLE LT Eh nmh Fa iamm CT Elution Technique lsocratic with Delayed Fractionation lsocratlc Length of Elution Before Fractionation 0 25 CV 0 25 CV Length of Elution with Fractionation 1 2 CV FIinish 4 9 Variables
188. mat 3 Use thick IIneS 4 Preview Customise Report Ex 5 OK 1 14 3 5 Customise Report Edit mode 1 Resultt 1 Open File Open Result 2 File Print Print ChromatogramS 3 Preview
189. of Gradient 3 Fraction Volume 1 ml FIinisSh 9 Variables Fe ee he Ee nl be en no II 1 FL E mi nn 1 1 PF TE Wizard Variables 62 Wavelength UV2 UV3 2 3 Value ON OFF Enter lt VariaDleSs gt Wavelength 2 3 Wash Out Unbound Sample Clean after Elution at 100 B Reequilibration PeakFrac Parameters Slope Slope VariableS Show detailsS Frac 950
190. ow it 18mm gt 15ml 12mm Violet 12mm 1 5ml 2mlmin 8 1 4 DropSyncro Accumulator DropSyncro 2mlmin Accumulator C mmin ml 18mm Yellow 18mm gt 15ml 12mm Violet 12mm l15m 96 well Plate Blue 96 S38 2ml min 8 1 4 DropSyncro Accumulator DropSyncro 2mlmin Accumulator 1 Greiner high 780201 Nunclow 156545 Greiner PP Masterblack 2ml 96well 780270 2
191. t 100 B ReeduilibratiOn PeakFrac Parameters Slope Slope VariableS Show detaillsS Frac 950 gt VariaDbleS Frac 950 Rack Wizard Fraction Order Serpentine row 4 VariaDleS BufferPreDp BufferPrep ON BufferPrep Recipe 2 3 0 7 5pH CIEX 11 SCOutindg Ea HE TL i Fmm Fae Euler fi Mgah eT a ded a Fw PP es kil Ca meee ed BufferPreD pH OK 12 EE Th uF 1T 1 i Em 1 mm i
192. tion Position 2 BufferPrebp OFF Next gt 3 Wavelength UV1 1 280nm 2 3 Variables 9 Pump A Inlet A1 Pump B Inlet B1 Buffer Valve A Inlet A11 Start Concentration BB 0 B Equilibration Volume 5 CV CV A 3 5CV Next gt 60 4 TL FCL eri eh Fein AN rm s miE rw 5 nm mrmim nm FmHh in HH nmem TI Injection techniqdue Sample Pump Direct Loading P 950 Injection Volume 100 ml Sample Inlet S1 S 8 Rinse Sample FlowPath 20 ml S8 OFF Flush Sample up to Injection Valve 10 ml
193. ube FirstTube 1 0 Segment FirstTube Start at FirstTube Variables Show detall lt Frac 900 gt Peak Identificatlon Level UV Level Slope Varables 9 Peak Fraction Volume ml Peak Fractionation ON Fixed Volume Fractionation
194. uuu 76 2 4 EETKKII Y jRWjaw x 3 81 O 0 TO TERKREIIYVYKWAYKAAOOWIWO IYI TTYTIKEIKEREUTVTKTKEKKYKTYYYT1TvTCKCLTO 86 86 2 ODM 2 ETEIITIKKIANKTKTKKTKRKKYTWII7xmwiIWT7irmuimiijmnmnmwsjnswnwwm 86 es 86 eo 86 12 1 2 kk 87 2 2 0 KK KK 55 87 12 3 pH kk 88 12 4 lt 89 13 90 13 1 90 13 2 kk 90 2 Rungalad Ni 90 13 2 2 CUNeS sal 91 ODODRGRGRGDOD Ai 92 kk 92 Y 93 X kk 93 94 13 2 3 kk 94 182 4 Lod OR 94 13 3 kk 95 eee 95 13 3 2 95 EE EE eo 95 14 Evaluation kk 96 14 1 kk 96 14 2 EEEEE EKk Eee 96 42 1 NN 97 14 2 2 97
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