ÄKTAFPLC UNICORN 5.0 71-1945-35
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
Contents 1....4 1.1 ÄKTAFPLC...4 1.2 ÄKTAFPLC...8 2....9 2.1 UNICORN...9 2.2 UNICORN...11 3....13 3.1...13 3.2...13 3.3 UV...13 3.4 PumpWash...14 3.5 SystemWash...14 4....15 4.1...15 4.2...16 4.3...16 4.4...17 5....18 5.1...18 5.2...19 6....20 6.1 Frac-950...20 6.2 Frac-900...24 7....28 7.1 Wizard...28 7.2...29 7.3 Equilibration...30 7.4...30 7.5...31 7.6 Elution...34 7.7 Segment 1...35 7.8 Segment 2...37 7.9 Variables...38 7.10 Run SetUp...39 7.11...40 7.12...40
8....41 8.1...41 8.2...41 8.3...42 8.4...42 8.5...43 8.6...43 8.7...44 8.8...49 9....50 9.1...50 9.2...50 9.3...53 10....54 10.1...54 10.2...54 10.3...54 10.4...56 10.5 UV...56 11....57 11.1...57 11.2...57 11.3...58 11.4...58 11.5...58 11.6...58 12....59 12.1...59 12.2 Copy to External...60 12.3 UNICORN CD...60 12.4 Copy from External...62 12.5...63 12.6 Rename...63
1 ÄKTAFPLC On-line Help 1.1 ÄKTAFPLC UNICORN P-920 ÄKTAFPLC 2 0.05 20 ml/min 5 MPa UPC-900 UV ph ph UV 280 254 nm Hg UV 0.01-5 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
ÄKTAFPLC 1 INV-907 UPC-900 M-925 500 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 1 7 6 2 3 4 5 G3 600 (350+250) Injection valve W2 1500 W2 W1 1500 W1 UV G4 170 G6 Cond Frac-900 = 330 Frac-920 = 550 Male/Male G5 600 Flow restrictor 0.2 MPa W3 1700 W3 Fraction collector 5
M-925 M-925 0.6 ml 2 ml 5 ml 12 ml 0, 6 V1 3 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 V1 2 6 100 µl 2 ml 150 ml 10 ml 50 ml 150 ml 3 FR- 902 0.2 MPa 2 µm 18-1027-11 10 6 Frac-900 95 10 18 mm 175 12 mm 40 30 mm 3 Frac-900 Frac-950 XY 4 A 120 18 & 30 mm B 240 12 mm C 96 4 30 mm 8 D 45 30 mm Frac-950 Extra rack panel for optional equipment 7
1.2 ÄKTAFPLC & 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 2 1 1 CU-950 PC 4 1. CU-950 2. ÄKTA ON PC OFF ON 3. ON Login Windows 4. Windows UNICORN 5. Logon User name default Password default 6. ÄKTA UNICORN Manager 9
6. ÄKTA System Control Run data Instruments Ready CU-950 LED Power PC System 3 um 10
2.2 UNICORN 2 2.2.1 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
2.2.2 1 2. 3. Close 4. 5. 5-1 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
3 3 System Control Manual 3.1 W1, W2, W3 3 3.2 20 57 18-1032-11 50cm 20 3.3 UV UV 30 ON 30 1. UPC-900 Lamp (on) 2. Lamp ON/OFF OK ON OK 3. 4. Hg 280nm 254nm Lamp (on) 280 nm 254 nm on off UPC-900 System Control System Settings Specials Keyboard Mode Open OK 13
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
4 4 4.1 ÄKTAdesign 1/16 HiTrap FPLC HR M6 2004 10 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 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 15
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 4.3 20% 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
4 HiPrep ÄKTA HiPrep 0.15MPa ÄKTA 0.35 MPa HiPrep 0.15 MPa FR-902 0.2 MPa HiTrap ÄKTA HiTrap 0.5 MPa HiTrap 0.3 MPa FR-902 0.2 MPa 4.4 1. 2. Manual Pump PumpWash ON, ON Execute 1 A 2 A/B 17
18 um 5.1 1. 2 6 2. 3 3. 4 5 0.7mm 90 6 2 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 2 6 18-1112-51 Superloop 150 ml 50 ml 10 ml Superloop 10 ml 50 ml 4 MPa Superloop 150 ml 2 MPa 19
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 20
6 6.1.2 Frac-950 2 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 2. 25 cm PEEK 5 mm C A B C 6.1.3 Keying guide Knurled screw 21
6.1.4 12mm 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 1 1 2 Greiner high 780201 Nunc low 156545 Greiner PP Masterblack 2ml 96well 780270 um 22
6 6.1.5 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 90/100 19.6 100µl 118µl 348 118 466µl Accumulator PEEK mm ) µl / 10 cm PEEK 0.5 19.6 ÄKTAFPLC 23
6.2 Frac-900 6.2.1 1 2 Pause Continue um 24
3 6 4 1 5 5 mm 6 PEEK 5 mm 25
7 PEEK 8 1 3 12 mm 30 mm 10-18 mm 12 mm Eppendorf tube holder for Tube Rack 175 12 mm 1.5 ml 0.5 ml 30 mm um 26
6 6.2.2 ph UV System Control System Settings... Specials FracParameters_900 ph ph Frac-900 0.5mm 375 µl 493 µl Frac-920 0.5mm 418 µl 536 µl ph PEEK ph 0.5mm 90mm 0.5mm 100mm 19.6µl ph 100µl ph 90/100 19.6 100µl 118µl 375 118 493µl PEEK mm ) µl / 10 cm PEEK 0.5 19.6 ÄKTAFPLC 27
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-900 7.1 Wizard Method Editor File New... New Method Wizard OK um 28
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
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 2 3 1.2 1 Next um 30
7.5 7 7.5.1 Frac-950 Frac-900 7.5.2 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
Elution Fractionation Frac-950 Frac-950 OFF 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 mm 30 mm Fraction Volume 1 ml Tube Type 18 mm Start at First Tube Next um 32
7 7.5.2 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-900 3 Fixed Volume and Peak Fractionation Fixed Volume Fractionation Peak Fractionation Fixed Volume Fractionation Fraction Volume 1 ml Next 33
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
7 7.7 Segment 1 1 2 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......10CV Next 35
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 7.5.1 7.5.2 Fraction Volume Wash Inlet A............................... Wash Inlet B............................... Flush System with Set Concentration B 5ml... Fraction Volume............................ 1ml Next um 36
7.8 Segment 2 7 Segment 2 1 Elution-Segment 2[2] --- Page 1[2]...................Step 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 37
7.9 Variables Wizard Variables Value Variables Wash Out Unbound Sample Reequilibration Flexible Flow Rates Variables Show details um 38
7.10 Run SetUp 7 Frac-950 Rack Fraction order Wizard 4 Gradient X min / ml / CV 39
7.11 File Save As... Method name Technique OK 001 Result 7.12 UNICORN Manager Method um 40
8 8 System Control 8.1 0.45 µm ph 8.2 1. 2 6 2. Load 3 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 3 4 41
8.3 Frac-950 6.1.1 Frac-900 8.4 1. System Control File Run... 2. OK 3. Next 4. Frac-950 Rack Fraction order 5. Start Next Start um 42
8.5 8 1. END End Method 2. save partial result OK 8.6 1. View Panes... 2. Rundata Curves OK 3. 43
8.7 8.7.1 Run Data 1. Run Data 2. Properties Run Data Groups 3. 4. New Group 5. OK um 44
8.7.2 Curves 8 1. Curves 2. Properties Curves 3. Select All 4. OK 1. Curves Marker 2. 3. X Y 4. X Y 5. Marker 45
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 46
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 1. 2. 3. 4. Undo Zoom 5. Reset Zoom 47
8.7.3 1. View Panes... 2. Flow Scheme 3. 4. 8.7.4 Logbook 1. View Panes... 2. Logbook 3. 4. Result um 48
8.8 8 System Control Run Hold Pause Continue End Continue Continue Pause Hold Pause 1. Manual Execute 49
9 9.1 1. Evaluation 2. File Open Result... 3. Open Result OK 9.2 1. 2. Properties 3. Chromatogram Layout 9.2.1 1. Chromatogram Layout Curves 2. 3. OK um 50
9.2.2 9 Y 1. Chromatogram Layout Y-Axis 2. 3. 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
9.2.3 1. 2. 3. 4. Page Down Page Up 5. 6. Reset zoom um 52
9 9.2.4 1. Add Text... 2. abc 3. abc 4. text text 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 53
10 2 20% 10.1 System Control 1. Manual Pump... PumpWash ON, ON Execute 10.2 1. 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-950 1. Manual Pump Flow 2ml/min Execute 2. Accumulator Accumulator Manual Frac AccumulatorWash Strokes 5 Execute 3. Accumulator END um 54
Frac-900 10 1. Manual Pump... Flow 2ml/min Execute 2. 3. Manual Frac... Fractionation_900 FracSize 2ml Execute 4. Manual Frac... Fractionation_Stop_900 Execute 5. END 55
10.4 1. 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-900 2. Lump ON/OFF OK OFF OK Lamp (on) Lamp (on) on off um 56
11 11 11.1 2 20% 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 11.2 20% 1. 2. 50%B 20ml/min 3. 4. 5. 6. 7. 8. 1-6 1 2 57
11.3 18-1027-11 10 11.4 11-0004-14 11.5 Frac-900 19-6067-02 11.6 18-1154-17 1. 2. 3. um 58
12 12 12.1 2HD DOS-V 1.44MB 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 59
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
4. My Computer CD UNICORN Result C:/UNICORN/Local/Fil CD C:/UNICORN/Local/Fil/Defaut /Result Manual Runs 12 5. CD Write these files to CD 6. CD Writing Wizard CD name Next 7. Finish 61
12.4 Copy from External 1. UNICORN Manager Methods Results 2. 3. Copy from External... 4. 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
12.5 12 1. UNICORN Manager Methods Results 2. Ctrl 3. Delete 4. OK 12.6 Rename 1. UNICORN Manager Methods Results 2. Rename 3. OK 63
ÄKTAFPLC Pause Continue No Yes 1 Manual Pump PumpWash ON, ON Execute 2 Manual Pump Flow 5 ml / min Execute END um 64
ÄKTAFPLC 0.00MPa 0.05MPa NO YES SystemControl System Calibrate... Monitor P920Press Start caribrate 65
ÄKTAFPLC 0.15 0.35MPa Yes No 0.15MPa Check 2 0.35MPa 0.10MPa No Yes Check 3 Act 1 um 66
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-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'
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