[PDF] AKTA FPLC UNICORN 5.0版

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1 ÄKTAFPLC UNICORN

2 UNICORN Ver.5.0 UNICORN Ver.5.0 PumpWash Continue PumpWash Pause Continue Frac-950 Wizard Elution Fractionation Start at First Tube Segment1 Fraction Volume Fraction Volume 0 First Tube Segment Segment1 First Tube OFF 10 ON UV Cond/pH Frac-900 Wizard Elution Technique Gradient Segmented Gradient Advanced Segmented Gradient Advanced

3 Contents ÄKTAFPLC ÄKTAFPLC UNICORN UNICORN UV PumpWash SystemWash Frac Frac Wizard Equilibration Elution Segment Segment Variables Run SetUp

4 UV Copy to External UNICORN CD Copy from External Rename...63

5 1 ÄKTAFPLC On-line Help 1.1 ÄKTAFPLC UNICORN P-920 ÄKTAFPLC ml/min 5 MPa UPC-900 UV ph ph UV nm Hg UV AU 313,365,405,463,546 nm 214 nm Zn 214 nm 5 mm 6 µl 2 mm 2 µl 2 mm ph flow cell with ph electrode mounted Conductivity flow cell with integrated temperature sensor UV flow cells with 2 and 5 mm path lengths um 4

6 ÄKTAFPLC 1 INV-907 UPC-900 M µl P-920 UV UV W1 W2 FR-902 A 1700 Mixer Pump A G1 600 Male/Male B 1900 Pump B On-line filter Stop plug G2 400 Loop 100 ml G3 600 ( ) Injection valve W W2 W W1 UV G4 170 G6 Cond Frac-900 = 330 Frac-920 = 550 Male/Male G5 600 Flow restrictor 0.2 MPa W W3 Fraction collector 5

7 M-925 M ml 2 ml 5 ml 12 ml 0, 6 V1 3 Load 3 Inject Waste PumpWash Column Sample syringe Pump Column Pump Column Pump Waste Load Waste Inject Waste Waste Waste um 6

8 1 V µl 2 ml 150 ml 10 ml 50 ml 150 ml 3 FR MPa 2 µm Frac mm mm mm 3 Frac-900 Frac-950 XY 4 A & 30 mm B mm C mm 8 D mm Frac-950 Extra rack panel for optional equipment 7

9 1.2 ÄKTAFPLC & 500 ml 20 % ÄKTA % Evaluation ÄKTA 9 10 um 8

10 UNICORN ÄKTA CU-950 PC 4 1. CU ÄKTA ON PC OFF ON 3. ON Login Windows 4. Windows UNICORN 5. Logon User name default Password default 6. ÄKTA UNICORN Manager 9

11 6. ÄKTA System Control Run data Instruments Ready CU-950 LED Power PC System 3 um 10

12 2.2 UNICORN UNCORN UNICORN 4 UNICORN Manager Method Editor System Control Evaluation 6 UNICORN Manager Copy, Rename, Delete, Move Method Editor System Control Evaluation Instant Run Logon/Logoff UNICORN Logon/Logoff Method Queue Method Queue Editor Display running Method Queue 11

13 Close Manual Manual 5-2 Pump... Manual Pump 5-3 Pump Wash Manual Pump PumpWash 5-4 Manual Pump PumpWash ON, ON 5-5 Execute um 12

14 3 3 System Control Manual 3.1 W1, W2, W cm UV UV 30 ON UPC-900 Lamp (on) 2. Lamp ON/OFF OK ON OK Hg 280nm 254nm Lamp (on) 280 nm 254 nm on off UPC-900 System Control System Settings Specials Keyboard Mode Open OK 13

15 3.4 PumpWash 2 20% 1. System Control 2. Manual Pump PumpWash ON, ON Execute 5 PumpWash Waste W1 PumpWash Continue Pause Continue 3.5 SystemWash 1 UV G3 1. Manual Pump Flow 5ml/min Execute UV, Cond, Pressure 5 End Load G3 UV W3 um 14

16 ÄKTAdesign 1/16 HiTrap FPLC HR M HiTrap HiTrap 1/16 PEEK ÄKTAFPLC 0.5 mm i.d. PEEK Tricorn RESOURCE 1/16 M6 1/16 ÄKTAdesign 1/16 M6 HiTrap HiLoad FPLC HR M6 Union M6 female / 1/16 male PEEK Union 1/16 female / M6 male PEEK Union 1/16 male / M6 female PEEK Union 1/16 male / Union 1/16 male PEEK Fingertight connector, 1/16 male PEEK Stop plug, 1/16 male PEEK Union, Luer female 1/16 male PEEK SRTC-2, M6 female (1.5 mm i.d.) PEEK Tubing (0.25 mm id, blue) PEEK 2 m Tubing (0.50 mm id, orange) PEEK 2 m Tubing (0.75 mm id, green) PEEK 2 m Tubing (1.0 mm id, beige) PEEK 2 m

17 4.2 HiTrap HiPrep UNICORN Manual Alarms&Mon... Alarm_Pressure High Alarm Execute 1 Manual Pump... Flow 0.5 ml/min Execute 2 UV 500 µl 0,6 l m 3 END % 1 Pressure limit Manual Alarms&Mon... Alarm_Pressure High Alarm Execute 2. 20% Manual Pump... Flow 1/2 Execute 1/4 3 3 UV Cond Pressure END um 16

18 4 HiPrep ÄKTA HiPrep 0.15MPa ÄKTA 0.35 MPa HiPrep 0.15 MPa FR MPa HiTrap ÄKTA HiTrap 0.5 MPa HiTrap 0.3 MPa FR MPa Manual Pump PumpWash ON, ON Execute 1 A 2 A/B 17

19 18 um mm Sample syringe Waste Column Waste Pump Inject Column Waste Pump Load Column Waste Waste Pump

20 Superloop 150 ml 50 ml 10 ml Superloop 10 ml 50 ml 4 MPa Superloop 150 ml 2 MPa 19

21 6 Frac-950 Frac Frac DropSync 2 ml/min Accumulator Accumulator 2 ml/min Accumulator Tube DropSync Accumulator OFF System Control System Settings... Specials FracParameters um 20

22 Frac ml/min DropSync Tube 1 G5 FR-902 Frac Accumulator A 2 5 mm C 3 Accumulator PEEK A 2 ml/min Accumulator 1. G5 Accumulator B cm PEEK 5 mm C A B C Keying guide Knurled screw 21

23 mm 18mm 30mm 96 30mm 18mm 30mm 2 ml/min DropSync 2 ml/min Accumulator Rack A 18mm 15 ml 1 Rack B 12mm 5 ml Rack C 96 2 Rack D 30mm 50 ml Greiner high Nunc low Greiner PP Masterblack 2ml 96well um 22

24 Accumulator ph UV System Control System Settings... Specials FracParameters Accumulator ph ph 0.5 mm 348 µl 466 µl 0.5 mm 197 µl 315 µl ph PEEK ph 0.5mm 90mm 0.5mm 100mm 19.6µl ph 100µl ph 90/ µl 118µl µl Accumulator PEEK mm ) µl / 10 cm PEEK ÄKTAFPLC 23

25 6.2 Frac Pause Continue um 24

26 mm 6 PEEK 5 mm 25

27 7 PEEK mm 30 mm mm 12 mm Eppendorf tube holder for Tube Rack mm 1.5 ml 0.5 ml 30 mm um 26

28 ph UV System Control System Settings... Specials FracParameters_900 ph ph Frac mm 375 µl 493 µl Frac mm 418 µl 536 µl ph PEEK ph 0.5mm 90mm 0.5mm 100mm 19.6µl ph 100µl ph 90/ µl 118µl µl PEEK mm ) µl / 10 cm PEEK ÄKTAFPLC 27

29 7 Wizard 1 Wizard Wizard RESOURCE S 1 ml 4 ml/min 100 µl 100 µl Segmented Gradient Advanced 50 %B 10 CV 280 nm Frac-950 Frac Wizard Method Editor File New... New Method Wizard OK um 28

30 7 7.2 Main Selection Affinity Anion Exchange Cation Exchange CIP HIC Installation test RPC Size Exclusion Cation Exchange Column RESOURCE S 1 ml Flexible Flow Rates 3 Variables Flow Regulation of the System Pump Flexible Flow Rate Next 29

31 7.3 Equilibration Start Concentration B B% 0 %B Equilibration Volume Watch Equilibration CV 3 5 CV Conductivity UV OFF Next 7.4 Injection Technique Manual Manual Empty Loop With Next um 30

32 Frac-950 Frac Frac-950 Setting / Flowthrough Fractionation Frac-950 Rack 18 & 30 mm Tube Wash Out unbound Sample [2 CV] 2 CV Variables 7.9 Flowthrough Fractionation Frac-950/OFF Frac-950 Fraction Volume Tube Type Start at Next tube First tube 18 mm 30 mm 1 Fraction Volume 50 ml Tube Type 30 mm Start at First Tube Next 31

33 Elution Fractionation Frac-950 Frac-950 OFF Frac-950 Frac Fixed Volume and Peak Fractionation Fixed Volume Fractionation Peak Fractionation Fixed Volume Fractionation Fraction Volume Tube Type Start at 18 mm 30 mm Fraction Volume 1 ml Tube Type 18 mm Start at First Tube Next um 32

34 Frac-900 Wash Out Unbound Sample Wash Out unbound Sample [2 CV] 2 CV Variables 7.9 Flowthrough Fractionation Frac-900/OFF Frac-900 Frac-900 Fraction Volume Next Elution Fractionation Frac-900 Frac-900 OFF Frac-900 Frac Fixed Volume and Peak Fractionation Fixed Volume Fractionation Peak Fractionation Fixed Volume Fractionation Fraction Volume 1 ml Next 33

35 7.6 Elution Elution Technique Isocratic Isocratic Isocratic with Delayed Fractionation Linear Gradient 1 Segmented Gradient Advanced 9 Step Gradient Segmented Gradient Basic 9 Gradient Segmented Gradient Advanced Gradient Segments 2 Reequilibration after Elution (5CV) 5 CV Next um 34

36 7 7.7 Segment Segment 1 1 Elution-Segment 1[2] --- Page 1[2] Step / Gradient Gradient Target Concentration B Segment %B 100% Length of Gradient Segment CV 20CV Gradient Target Concentration B...50% Length of Gradient CV Next 35

37 Segment 1 2 Elution-Segment 1[2] --- Page 2[2] Segment 1 Wash Flush Wash Flush [Segment 1] Wash Inlet A A PumpWash 1 Wash Inlet B B PumpWash 1 Flush System with Set Concentration B 5ml A,B Step 1 ConcB% Fraction Volume Segment Fraction Volume Wash Inlet A Wash Inlet B Flush System with Set Concentration B 5ml... Fraction Volume ml Next um 36

38 7.8 Segment 2 7 Segment 2 1 Elution-Segment 2[2] --- Page 1[2] Step Target Concentration B...100% Length of Gradient CV Next Segment 2 2 Elution-Segment 2[2] --- Page 2[2] Wash Inlet A Wash Inlet B Flush System with Set Concentration B 5ml... Fraction Volume ml Finish 37

39 7.9 Variables Wizard Variables Value Variables Wash Out Unbound Sample Reequilibration Flexible Flow Rates Variables Show details um 38

40 7.10 Run SetUp 7 Frac-950 Rack Fraction order Wizard 4 Gradient X min / ml / CV 39

41 7.11 File Save As... Method name Technique OK 001 Result 7.12 UNICORN Manager Method um 40

42 8 8 System Control µm ph Load 3 3 Column Sample syringe Pump Column Pump Column Pump Waste Load Waste Inject Waste Waste Waste

43 8.3 Frac Frac System Control File Run OK 3. Next 4. Frac-950 Rack Fraction order 5. Start Next Start um 42

44 END End Method 2. save partial result OK View Panes Rundata Curves OK 3. 43

45 Run Data 1. Run Data 2. Properties Run Data Groups New Group 5. OK um 44

46 8.7.2 Curves 8 1. Curves 2. Properties Curves 3. Select All 4. OK 1. Curves Marker X Y 4. X Y 5. Marker 45

47 1. Curves Properties 2. Curve Style and Color OK Y Y 1. Curves Properties 2. Y-Axis 3. Fixed OK um 46

48 X 8 X Time min Volume ml 1. Curves Properties 2. X-Axis 3. Time min Volume ml 4. Total Window Total Window 5. OK Undo Zoom 5. Reset Zoom 47

49 View Panes Flow Scheme Logbook 1. View Panes Logbook Result um 48

50 8.8 8 System Control Run Hold Pause Continue End Continue Continue Pause Hold Pause 1. Manual Execute 49

51 Evaluation 2. File Open Result Open Result OK Properties 3. Chromatogram Layout Chromatogram Layout Curves OK um 50

52 Y 1. Chromatogram Layout Y-Axis Auto Fixed 4. OK X 1. Chromatogram Layout X-Axis 2. Auto Fixed 3. Adjust retention zero to injection number 0 min ml 4. OK 51

53 Page Down Page Up Reset zoom um 52

54 Add Text abc 3. abc 4. text text Edit Text Mode File Print... Print Chromatograms 2. Print Format 3. Landscape 4. Preview... Customise Report Exit 5. OK 53

55 % 10.1 System Control 1. Manual Pump... PumpWash ON, ON Execute Pressure limit Manual Alarms&Mon Alarm_Pressure High Alarm Execute 2. Manual Pump Flow Execute 3. 3 UV, Cond, Pressure END 4. UV G3 5. Manual Flowpath Injection Valve Inject Execute Manual Pump Flow 5ml/min Execute 6. 5 END 10.3 Frac Manual Pump Flow 2ml/min Execute 2. Accumulator Accumulator Manual Frac AccumulatorWash Strokes 5 Execute 3. Accumulator END um 54

56 Frac Manual Pump... Flow 2ml/min Execute Manual Frac... Fractionation_900 FracSize 2ml Execute 4. Manual Frac... Fractionation_Stop_900 Execute 5. END 55

57 UNICORN Manager 2. File Quit program 3. Unicorn Manager Yes 4. System Control Leave Control of System1 OK 5. Windows Start Turn off Computer Turn Off ÄKTAexplorer 6. OFF OFF 10.5 UV UV OFF 1. UPC Lump ON/OFF OK OFF OK Lamp (on) Lamp (on) on off um 56

58 % 20% 20% ph ph 1. 20% 2. System Control Manual Pump... PumpWash ON, ON Execute 3. PumpWash Manual Pump... Flow 5ml/min Execute 4. 5 END % %B 20ml/min

59 Frac um 58

60 HD DOS-V 1.44MB 1. A 2. Windows My Computer /2 Floppy (A:) 4. Format Start 6. OK 7. Format complete OK 8. Close 9. My Computer 59

61 12.2 Copy to External 1. UNICORN Manager Methods Results 2. Copy to External 3. Copy to: 3 1/2 Floppy (A:) Save Copy to external Result zip zip Copy from External 12.3 UNICORN CD 1. CD CD 2. OK 3. CD Delete temporary files um 60

62 4. My Computer CD UNICORN Result C:/UNICORN/Local/Fil CD C:/UNICORN/Local/Fil/Defaut /Result Manual Runs CD Write these files to CD 6. CD Writing Wizard CD name Next 7. Finish 61

63 12.4 Copy from External 1. UNICORN Manager Methods Results Copy from External Copy: 3 1/2 Floppy (A:) 5. OK 6. Result Results 7. Method Method-System Connection 8. Method file Systems System 1 9. OK Copy from external zip Copy zip um 62

64 UNICORN Manager Methods Results 2. Ctrl 3. Delete 4. OK 12.6 Rename 1. UNICORN Manager Methods Results 2. Rename 3. OK 63

65 ÄKTAFPLC Pause Continue No Yes 1 Manual Pump PumpWash ON, ON Execute 2 Manual Pump Flow 5 ml / min Execute END um 64

66 ÄKTAFPLC 0.00MPa 0.05MPa NO YES SystemControl System Calibrate... Monitor P920Press Start caribrate 65

67 ÄKTAFPLC MPa Yes No 0.15MPa Check MPa 0.10MPa No Yes Check 3 Act 1 um 66

68 CU-950 CU-950 CU-950 ÄKTA 1 ÄKTA 1. Uninet ÄKTA CU-950 Uninet 2 ÄKTA CU USB CU-950 USB USB USB 3. CU-950 CU CU-950 Power PC System ÄKTA 4. ÄKTA ÄKTA Windows CU-950 Power PC System PC 2 5. UNICORN ÄKTA UNICORN ÄKTA CU-950 Power PC System PC CU-950 USB CU-950 System Uninet CU-950 'Test' 'Network' 'UniNet-1' Input 24-32V 1A USB UniNet-1 Test 3 Fraction Collector Frac-920 Frac-920 Frac-900 Frac-900 Frac-920 ÄKTA Setup and Check ÄKTA UNICORN Never 'Properties' Display Properties Screen Saver 'Power...' Power Options Properties Plugged in System standby 'Never'

69 60

70 OFF TEL : : : 30

71 TEL FAX MicroCal Q&A Science Program Web GE 8

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