ÄKTApurifier UNICORN 5.0 71-1944-35
UNICORN Ver.5.0 UNICORN Ver.5.0 PumpWash PurifierContinue PumpWash Pause Continue Frac-950 Frac-950 2 Manual Flowpath OutletValve; F2 Execute Manual Frac Man_Fractionation Execute Frac-950 WizardElution Fractionation Start at First Tube Segment1 Fraction Volume Fraction Volume 0 First Tube Segment Segment1First Tube OFF10ON UV Cond/pH Frac-900 WizardElution Technique Gradient Segmented Gradient Advanced Segmented Gradient Advanced
Contents 1....4 1.1 ÄKTApurifier...4 1.2 ÄKTApurifier...8 2....9 2.1 UNICORN...9 2.2 UNICORN...11 3....13 3.1...13 3.2...13 3.3...14 3.4...15 3.5 PumpWash...15 3.6...16 4....17 4.1...17 4.2...18 4.3...18 4.4...19 5....20 5.1...20 5.2...21 6....22 6.1 Frac-950...22 6.2 Frac-900...26 7....30 7.1 Wizard...30 7.2...31 7.3...32 7.4 Equilibration...32 7.5...33 7.6...33 7.7 Elution...36 7.8 Segment 1...37 7.9 Segment 2...39 7.10 Variables...40
7.11 Run Setup...41 7.12...42 7.13...42 8....43 8.1...43 8.2...43 8.3...44 8.4...44 8.5...45 8.6...45 8.7...46 8.8...51 9....52 9.1...52 9.2...53 9.3...56 10....57 10.1...57 10.2...57 10.3...57 10.4...59 11....60 11.1...60 11.2...60 11.3...61 11.4...61 11.5...61 11.6...61 12....62 12.1...62 12.2 Copy to External...63 12.3 UNICORN CD...63 12.4 Copy from External...65 12.5...66 12.6 Rename...66
PV-908 INV-908 1 ÄKTApurifier On-line Help 1.1 ÄKTApurifier Box-900 INV-907 ph/c-900 UV-900 PV-908 M-925 P-903 M-925 (W1, W2 and W3) A SV-903 B SV-903 UV G1 Pump B A3 G3 Pump A G4 On-line filter Mixer G2 B3 New capillary of 180 mm ph Flow cell G5 (Bypass) Injection Valve (V1) 1 2 7 6 3 4 5 Column Fraction Collector NC NO NC NO Sample loop A2 A1 B2 B1 W2 W1 8 1 7 W3 2 6 3 Outlet Valve (V4) 4 5 G6 (H6) COND UV G8 (H8) New capillary of 130 mm Flow restrictor G9 (H9) ÄKTApurifier F3 um 4
P-903 1 2 0.001 10ml/min25MPa UV-900 190-700nmUV-Vis3 10mm 8µl 2mm 2µl ph/c-900 ph 5
6 Frac-900 9510 18 mm 17512 mm 4030 mm 3 Frac-950 XY4 A 12018 & 30 mm B 24012 mm C 96 430 mm 8 D 4530 mm Frac-900 Frac-950 V1 37 Load 3 Inject Waste PumpWash Column Sample syringe 2 4 1 3 5 7 6 Pump Column 2 4 1 3 5 7 6 Pump Column 1 2 3 4 5 7 6 Pump Waste Load Waste Inject Waste Waste Waste um 6
1 V126 10 µl 100 µl 1 ml 150 ml 10 ml 50 ml 150 ml 3 FR-902 0.2 MPaFR-904 0.4 MPa2 FR-902 FR-902 FR- 904 2 µm 2M NaOH 18-1120-94 2 67 11.3 7
1.2 ÄKTApurifier & 500 ml 20 % ÄKTA 2 3 5 4 4 6 20 % 7 8 10 10 Evaluation ÄKTA 9 10 um 8
2 2 2.1 UNICORN ÄKTA 21 1 CU-950 PC4 1. CU-950 2. ÄKTA ONPC OFF ON 3. ON Login Windows 4. Windows UNICORN 5. Logon User name default Passworddefault 6. ÄKTA UNICORN Manager 9
6. ÄKTASystem Control Run data Instruments Ready CU-950 LEDPower PC System 3 um 10
2.2 UNICORN 2 2.2.1 UNCORN UNICORN 4UNICORN 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
2.2.2 1 2. 3. Close 4. 5. 5-1 Manual Manual 5-2 Flowpath Manual Flowpath... 5-3 Pump A Inlet Manual Flowpath... Pump A Inlet 5-4 A2 Manual Flowpath... Pump A Inlet A2 5-5 Execute um 12
3 3 System ControlManual 3.1 W1 5 W2 4 W3 1F3 PEEK4 F3 F2 F3 W3 3.2 20% A B 18-1132-75 13
3.3 2 1 2 A1, A2, B1, B2 4 1 4A1, B1 2 A 3 4 10 ml 5 A 2 4 A1 6B2 7 A2, B2 Manual Flowpath... PumpAInlet A2 Execute A1A2 Manual Flowpath... PumpAInlet B2 Execute B1B2 8 2 4 A2, B2 1 A1 um 14
3.4 3 UV 1 System Control Run Data Curves 2 AA1 Manual Pump... Flow 5 ml/min Execute Pressure 3 BB1 Manual Pump... Gradient Target 100 %B, Length 0 min Execute Pressure 4 BB2 Manual Flowpath... PumpBInlet B2 Execute 5 AA2 Manual Pump... Gradient Target 0 %B Length 0 min Execute Manual Flowpath... PumpAInlet A2 Execute 6 End A1, A2, B1, B20.05MPa 3.5 PumpWash 1 System Control 25 Manual Pump... PumpWash Purifier ON, ON, ON, ON Execute PumpWashContinue Pause Continue PumpWash wastew1 15
3.6 1. UV G5 2. Manual Pump... Flow 5ml/min Execute 5UV Cond Pressure 3. END LoadG5 UV ph W3 um 16
4 4 4.1 ÄKTAdesign1/16HiTrap FPLC HR M6 2004 10 HiTrap HiTrap1/16 PEEKÄKTApurifier0.5 mm i.d. PEEK0.25 mm i.d. PEEK TricornRESOURCE1/16 M6 1/16 1/16 M6 ÄKTAdesign HiTrapHiLoad FPLC HRM6 Union M6 female / 1/16 male PEEK 5 18-3858-01 Union 1/16 female / M6 male PEEK 6 18-1112-57 Union 1/16 male / M6 female PEEK 8 18-1112-58 Union 1/16 male / Union 1/16 male PEEK 2 18-1120-93 Fingertight connector, 1/16 male PEEK 10 18-1112-55 Stop plug, 1/16 male PEEK 5 18-1112-52 Union, Luer female 1/16 male PEEK 2 18-1112-51 SRTC-2, M6 female (1.5 mm i.d.) PEEK 5 19-2143-01 Tubing (0.25 mm id, blue) PEEK 2 m 18-1120-95 Tubing (0.50 mm id, orange) PEEK 2 m 18-1113-68 Tubing (0.75 mm id, green) PEEK 2 m 18-1112-53 Tubing (1.0 mm id, beige) PEEK 2 m 18-1115-83 17
PV-908 INV-908 4.2 HiTrap HiPrep UNICORN Manual Alarms&Mon... Alarm_Pressure High Alarm Execute 1 Manual Pump... Flow 0.5 ml/min Execute 2 UV 3 END 4.3 20% 1 Pressure limit Manual Alarms&Mon... Alarm_Pressure High Alarm Execute 2. 20% Manual Pump... Flow 1/2 Execute 1/4 33UV Cond Pressure END um 18
4 HiPrep ÄKTA HiPrep 0.15MPa ÄKTA 0.35 MPaHiPrep0.15 MPa FR-902 0.2 MPa HiTrap ÄKTA HiTrap 0.5 MPaHiTrap0.3 MPa FR-9020.2 MPa 4.4 1. Manual Pump... Pumpwash Purifier ON, OFF, ON, OFF Execute 1 A1 2 A1/B1 BufferPrep A1 Buffer A2 Acid/Base B1 Water B2 Salt 19
20 um 5.1 1. 2 6 2. 3 3. 4 5 0.7mm 90 3 5 Sample syringe 1 2 3 4 5 6 7 Waste Column Waste Pump 1 2 3 4 5 6 7 Inject Column Waste Pump 1 2 3 4 5 6 7 Load Column Waste Waste Pump
5.2 5 26 1 2 7 3 6 4 5 18-1112-51 Superloop 150 ml 50 ml 10 ml Superloop 10 ml 50 ml 4 MPa Superloop 150 ml 2 MPa 21
6 6 Frac-950 Frac-900 6.1 Frac-950 6.1.1 DropSync 2 ml/min Accumulator accumulator 2 ml/min Accumulator Tube DropSync Accumulator OFF System Control System Settings... Specials FracParameters um 22
6 6.1.2 V4Frac-950 2 ml/min DropSync Tube 1 OutletValve F2 Accumulator A 25 mmc 3 AccumulatorPEEK A 2 ml/min Accumulator 1. OutletValve F2 Accumulator B 2. 25 cm PEEK 5 mmc A B C 6.1.3 Keying guide Knurled screw 23
6 6.1.4 12mm 18mm 30mm 96 30mm 18mm 30mm 2 ml/mindropsync2 ml/minaccumulator Rack A 18mm 15 ml 1 Rack B 12mm 5 ml Rack C 96 2 Rack D 30mm 50 ml 1 1 2 Greiner high 780201 Nunc low 156545 Greiner PP Masterblack 2ml 96well 780270 um 24
6 6.1.5 AccumulatorpH System Control System Settings... Specials FracParameters Accumulator ph Cell ph Cell 0.25 mm 238 µl 0.5 mm 382 µl 527 µl 0.25 mm 88 µl 0.5 mm 232 µl 354 µl PEEK ph Cell PEEK mm ) µl / 10 cm PEEK 0.25 4.91 PEEK 0.5 19.6 ÄKTApurifier 25
6 6.2 Frac-900 6.2.1 1 2 Pause Continue um 26
3 6 4 1 55 mm 6 PEEK5 mm 27
7PEEK 8 1 330 mm 10-18 mm 12 mm Eppendorf tube holder for Tube Rack 175 12 mm 1.5 ml 0.5 ml 30 mm um 28
6 6.2.2 ph System Control System Settings... Specials FracParameters_900 ph Cell ph Cell 0.25mm 88 µl 0.5mm 232 µl 354 µl PEEK ph Cell PEEK mm ) µl / 10 cm PEEK 0.25 4.91 PEEK 0.5 19.6 ÄKTApurifier 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 254 nm Frac-950 Frac-900 7.1 Wizard Method Editor File New... New Method Wizard OK um 30
7 7.2 Main Selection Affinity Anion Exchange Cation Exchange Chromatofocusing 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 BufferPrep phon / OFF Next 31
7.3 WaveLength UV1 WaveLength UV2 WaveLength UV3 Pump A Inlet 1 1 2 3 A A1 Pump B Inlet B B1 Next 280 nm 254 nm 215 nm 7.4 Equilibration Start Concentration B B% 0 %B Equilibration Volume Watch Equilibration CV 3 5 CV Conductivity, ph, UV OFF Next um 32
7 7.5 Injection Technique Manual System Pump Direct Loading Manual Empty Loop With 2 3 1.2 1 Next 7.6 7.6.1 Frac-950 Frac-900 7.6.2 Frac-950 Setting / Flowthrough Fractionation Frac-950 Rack 18 & 30 mm Tube Wash Out unbound Sample [2 CV] 2 CV Variables 7.10 Flowthrough Fractionation Frac-950/OutletValveF3/OFF Frac-950 33
Fraction Volume Tube Type Start at Next tube First tube 18 mm30 mm 1 Fraction Volume 50 ml Tube Type 30 mm Start at First Tube Next Elution Fractionation Frac-950 OFF OutletValve Frac-950 OutletValve Frac-950 Frac-950 3 Fixed Volume and Peak Fractionation Fixed Volume Fractionation Peak Fractionation Fixed Volume Fractionation Fraction Volume Tube Type Start at 18 mm30 mm Fraction Volume 1 ml Tube Type 18 mm Start at First Tube Next um 34
7 7.6.2 Frac-900 Wash Out Unbound Sample Wash Out unbound Sample [2 CV] 2 CV Variables 7.10 Flowthrough Fractionation Frac-900/OFF/OutletValveF3 OutletValveF3 Frac-900 Fraction Volume Next Elution Fractionation Frac-900 Frac-900 OFF OutletValve OutletValve Frac-900 Frac-900 3 Fixed Volume and Peak Fractionation Fixed Volume Fractionation Peak Fractionation Fixed Volume Fractionation Fraction Volume 1 ml Next 35
7.7 Elution Elution Technique Isocratic Isocratic Isocratic with Delayed Fractionation Linear Gradient 1 Segmented Gradient Advanced 9 Step Gradient StepWize Gradient Segmented Gradient Basic 9 Gradient Segmented Gradient Advanced Gradient Segments 2 Reequilibration after Elution (5CV) 5 CV Next um 36
7 7.8 Segment 1 12 Segment 1 1 Elution-Segment 1[2] --- Page 1[2] Step / Gradient Gradient Pump Inlet A and B A B A1 B1 Target Concentration B Segment%B100% Length of Gradient Segment CV20CV...................Gradient Pump Inlet A and B..... A1,B1 Buffer Valve A Inlet...... A11 Target Concentration B...50% Length of Gradient......10CV Next Segment 1 2 Elution-Segment 1[2] --- Page 2[2] Segment 1Wash Flush Wash Flush [Segment 1] Wash Inlet A APumpWash 1 37
Wash Inlet B BPumpWash 1 Flush System with Set Concentration B 5ml A,B 1 ConcB% Fraction Volume Segment 7.6.1 7.6.2 Fraction Volume Wash Inlet A............................... Wash Inlet B............................... Flush System with Set Concentration B 5ml... Fraction Volume............................ 1ml Next um 38
7.9 Segment 2 7 Segment 2 1 Elution-Segment 2[2] --- Page 1[2]...................Step Pump Inlet A and B...... A1,B1 Target Concentration B...100% Length of Gradient.......3CV 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............................ 1ml Finish 39
7.10 Variables Wizard VariablesValue Variables Wash Out Unbound Sample Reequilibration Flexible Flow Rates Variables Show details um 40
7.11 Run Setup 7 Frac-950 Rack Fraction order Wizard 4 Gradient Xmin / ml / CV 41
7.12 File Save As... Method Name Technique OK 001Result 7.13 UNICORN Manager Method um 42
8 8 System Control 8.1 0.45 µm ph 8.2 1. 2 6 3 2. Load3 3 Column Sample syringe 2 1 3 7 6 Pump Column 2 1 3 7 6 Pump Column 2 3 1 7 6 Pump 4 5 4 5 4 5 Waste Load Waste Inject Waste Waste Waste 3. 3 4. 4 34 43
8.3 Frac-950 6.1.1Frac-900 Outletvalve F3F3 8.4 1. System Control File Run... 2. OK 3. Next 4. Frac-950Rack Fraction order 5. Start Next Start um 44
8.5 8 1. ENDEnd Method 2. save partial resultok 8.6 1. View Panes... 2. Rundata Curves OK 3. 45
8.7 8.7.1 Run Data 1. Run Data 2. Properties Run Data Groups 3. 4. New Group 5. OK um 46
8.7.2 Curves 8 1. Curves 2. Properties Curves 3. Select All 4. OK 1. Curves Marker 2. 3. XY 4. XY 5. Marker 47
1. Curves Properties 2. Curve Style and Color 3. 4. OK Y Y 1. Curves Properties 2. Y-Axis 3. Fixed 4. 5. OK um 48
X 8 XTime minvolume ml 1. Curves Properties 2. X-Axis 3. Time minvolume ml 4. Total WindowTotal Window 5. OK 1. 2. 3. 4. Undo Zoom 5. Reset Zoom 49
8.7.3 1. View Panes... 2. Flow Scheme 3. 4. 8.7.4 Logbook 1. View Panes... 2. Logbook 3. 4. Result um 50
8.8 8 System Control Run Hold Pause Continue End Continue Continue Pause Hold Pause 1. Manual Execute 51
9 9.1 1. Evaluation 2. File Open Result... 3. Open Result OK um 52
9.2 9 1. 2. Properties 3. Chromatogram Layout 9.2.1 1. Chromatogram Layout Curves 2. 3. OK 53
9.2.2 Y 1. Chromatogram Layout Y-Axis 2. 3. AutoFixed 4. 3UVAll with this unit 5. OK X 1. Chromatogram Layout X-Axis 2. Auto Fixed 3. Adjust retention zero to injection number 0 min ml 4. OK um 54
9.2.3 9 1. 2. 3. 4. Page Down Page Up 5. 6. Reset zoom 55
9.2.4 1. Add Text... 2. abc 3. abc 4. texttext 5. 1. Edit Text Mode 2. 3. 9.3 1. File Print... Print Chromatograms 2. Print Format 3. Landscape 4. Preview...Customise Report Exit 5. OK um 56
10 10 220% 10.1 System Control 1. Manual Pump... PumpWash Purifier ON, OFF, ON, OFF Execute 10.2 1. Pressure limit Manual Alarms&Mon Alarm_Pressure High Alarm Execute 2. Manual Pump Flow Execute 3. 3UV, Cond, Pressure END 4. UVG5 5. Manual Flowpath Injection Valve Inject Execute Manual Pump Flow 5ml/min Execute 6. 5END 10.3 Frac-950 1. Manual Pump Flow 2ml/min Execute 2. Manual Flowpath OutletValve F2 Execute 3. Accumulator Accumulator Manual Frac AccumulatorWash Strokes 5 Execute 4. Accumulator Manual Flowpath OutletValve F3 Execute 5. 3END 57
Frac-900 1. Manual Pump Flow 2ml/min Execute 2. 3. Manual Frac Fractionation_900 FracSize 2ml Execute 4. Manual Frac Fractionation_Stop_900 Execute 5. Manual Flowpath OutletValve F3 Execute 6. 3END um 58
10.4 10 1. UNICORN Manager 2. File Quit program 3. UNICORN Manager YES 4. System ControlLeave Control of System1OK 5. Windows Start Turn off Computer Turn off ÄKTAexplorer 6. OFF OFF 59
11 11.1 220%20% 20% ph ph 1. 20% 2. System Control Manual Pump... PumpWash Purifier ON, OFF, ON, OFF Execute 3. Pump Wash Manual Pump... Flow 5ml/min Execute 4. 5END 11.2 1. 20% 2. 3. 4. 13 um 60
11.3 11 2 µm 1 ml / min 0.35 MPa2M NaOH 18-1120-94 2 11.4 11-0004-14 1.5 Frac-900 19-6067-02 11.6 18-1154-17 1. 2. 3. 61
12 12.1 2HDDOS-V 1.44MB 1. A 2. WindowsMy Computer 3. 3 1/2 Floppy (A:) 4. Format... 5. Start 6. OK 7. Format completeok 8. Close 9. My Computer um 62
12.2 Copy to External 12 1. UNICORN Manager Methods Results 2. Copy to External 3. Copy 3 1/2 Floppy (A:) Save Copy to External Result zipzip Copy from External 12.3 UNICORN CD 1. CD CD 2. OK 3. CD Delete temporary files 63
4. My Computer CDUNICORN Result C:/UNICORN/Local/Fil CD C:/UNICORN/Local/Fil/Defaut /Result Manual Runs 5. CD Write these files to CD 6. CD Writing Wizard CD namenext 7. Finish um 64
12.4 Copy from External 12 1. UNICORN Manager Methods Results 2. 3. Copy from External... 4. Copy: 3 1/2 Floppy (A:) 5. OK 6. ResultResults 7. MethodMethod-System Connection 8. Method filesystems system 1 9. OK Copy from ExternalzipCopyzip 65
12.5 1. UNICORN Manager Methods Results 2. Ctrl 3. Delete 4. OK 12.6 Rename 1. UNICORN Manager Methods Results 2. Rename 3. OK um 66
ÄKTApurifier Pause Continue No Yes Check 2 END 67
ÄKTApurifier A2 0.00MPa 0.05MPa No Yes um 68
ÄKTApurifier Gradient ; ConcB% 0% Flow ml/min Execute 0.15 0.35MPa Yes No 0.15MPa Check 2 0.35MPa Gradient; Conc%B 0% InjectionValve Waste Execute 0.10MPa No Yes Check 3 Act 1 69
CU-950 CU-950 CU-950 ÄKTA 1 ÄKTA 1. Uninet ÄKTA CU-950 Uninet 2 ÄKTA CU-950 1 2. USB CU-950 USB USB USB 3. CU-950 CU-950 3 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-901 900 Frac-901 900 Frac-920 ÄKTA Setup and Check ÄKTA UNICORN Never 'Properties' Display Properties Screen Saver 'Power...' Power Options Properties Plugged in System standby 'Never'
UPC-900 ÄKTApurifier UPC 10/100 1. 280 nm 254 nm 2. Hg 280 nm 254 nm UV OFF 1. UPC-900 2. Lamp ON/OFF OK OFF OK Lamp (on) Lamp (on) on off UPC-900 System Control System Settings Specials Keyboard Mode Open OK memo
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