ÄKTA system Training Guide

ÄKTA system Training Guide
Explorer 100 / 10
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ÄKTAexplorer
ÄKTAexplorer
Injector
Buffers
Box
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ham
amers
Column
Valves
Monitor
Conductivity
pH
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amers
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bio scie
Outlet
Valve
UV Detector
3 Wavelengths
amersham bioscience
Pumps
am
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bios
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ence
ÄKTAexplorer
Pump A
pH/cell
electrode
Buffer switching
valve
Sample
Valve
Sample
holder
ÄKTAexplorer 100
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45
Mixer
System
pumps
Column
selection
valve (V2)
UV
3 Wavelength
detector
Cond
Conductivity
monitor
pH
A
1 87
2
6
345
B
Sample
pump
1 87
2
6
345
1 7
6
2
3 45
Reverse
flow valve (V7)
pH detector
Fraction
collector
Frac-950
1 87
2
6
345
1 87
2
6
345
Buffer
select
valve (V6)
Outlet
valve (V4)
1 87
2
6
345
4
Sample selection
valve (V5)
Column
selection
valve (V3)
ÄKTAexplorer 100 Air
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45
Mixer
System
pumps
Reverse
flow valve (V7)
Column
selection
valve (V2)
UV
3 Wavelength
detector
Cond
Conductivity
monitor
pH
A
1 87
2
6
345
B
Air
sensor
Sample
pump
1 87
2
6
345
1 7
6
2
3 45
pH detector
Fraction
collector
Frac-950
1 87
2
6
345
1 87
2
6
345
Buffer
select
valve (V6)
1 87
2
6
345
4
Sample selection
valve (V5)
Column
selection
valve (V3)
Outlet
valve (V4)
ÄKTAexplorer 10
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45
Mixer
System
pumps
Column
selection
valve (V2)
UV
3 Wavelength
detector
Cond
Conductivity
monitor
pH
A
1 87
2
6
345
B
pH detector
Fraction
collector
Frac-950
1 87
2
6
345
1 87
2
6
345
Buffer
select
valve (V6)
Outlet
valve (V4)
1 87
2
6
345
4
Column
selection
valve (V3)
ÄKTAexplorer 10 S
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45
Mixer
System
pumps
3 Wavelength
detector
Cond
Conductivity
monitor
pH
A
1 87
2
6
345
B
Sample
pump
1 87
2
6
345
Column
selection
valve (V2)
UV
pH detector
Fraction
collector
Frac-950
1 87
2
6
345
1 87
2
6
345
Buffer
select
valve (V6)
1 87
2
6
345
4
Sample selection
valve (V5)
Column
selection
valve (V3)
Outlet
valve (V4)
ÄKTAexplorer 10 XT
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45
Mixer
System
pumps
Column
selection
valve (V2)
UV
3 Wavelength
detector
Cond
Conductivity
monitor
pH
A
B
1 87
2
6
345
pH detector
Fraction
collector
Frac-950
1 87
2
6
345
Buffer
select
valve (V6)
1 87
2
6
345
1 87
2
6
345
4
Column
selection
valve (V3)
Outlet
valve (V4)
PUMP P-900
Mixer
Pump A
Pump B
Proportion
Valve
A2
Acid /Alkali
Ä KTA 10 mls/min
Ä KTA 100 mls/min
A1 B2
Buffer
B1
Salt
GRADIENT
<6 mls/min
6 -14 mls/min
<30 mls/min
30 - 50 mls/min
50 - 100 mls/min
Water
MIXER
0.6 ml mixer
2 ml mixer
2 mls mixer
5 mls mixer
12 mls mixer
PUMP P-900 Inlets
Mixer
Pump B
Pump A
Switching
Valves
Buffer Select
Valve IV-908
A2
A1
2
3
A11
A12
A13
1 8
4
A14
B1
Water
7
5
6
A16
A15
B2
A17
A18
Salt
PUMP P-900 Manually Priming Buffer Inlet Lines
Pump A
Common inlet
A2
Pump B
A1
Common inlet
Manual
Flowpath
Manual
A1 or A2
BufferValveA B1 or B2
B1
B2
Flowpath
BufferValveB
Open
Draw A
ir
out of
Buffer
Line
PUMP P-900 Back - Wash Lines
Pump Back-Wash
Lines
Syringe fitting
Wash OUT
IN-LINE
one-way
valve
Buffer
OUT
Ball outlet
check valve
High pressure
Piston seal
Piston
20% Ethanol : Water
Backwash pump
diaphragm
Ball inlet
check valve
Wash
IN
Buffer
IN
ON-LINE FILTERS
P-903 ( 10 ml/min Pump )
Low Flow HPLC Filter
P-901 ( 100 ml/min Pump )
High Flow Fibre Filter
Polymer Support
LOAD (sample)
INJECT (sample)
Column
Column
Column
2
3
2
Pump
7
5
8
Sample
7
4
Waste
4
5
Sample
6
3
6
3
6
8
2
2
5
5
PV-908
1
8
7
7
6
4
PV-908
1
2
4
5
Pump
7
3
6
4
PV-908
3
2
Pump
7
3
6
4
1
1
1
1
LOAD (wash loop)
Waste
5
Sample
Waste
INJECTION VALVE INV- 907
1
COLUMN
7
2
LOAD
SAMPLE
3
PUMP
6
4
5
Waste
Waste
LOAD
1
COLUMN
7
2
PUMP
3
4
6
5
Waste
Waste
INJECT
1
COLUMN
7
2
PUMP
3
4
Waste
6
5
Waste
WASTE
SAMPLE LOADING METHOD
1
COLUMN
2
LOAD
SAMPLE
7
PUMP
3
6
5
4
Needle Port
Waste
Waste
Always tighten by hand
with needle inserted
Do Not Overtighten
1
COLUMN
LOAD
SAMPLE
7
2
PUMP
3
6
4
5
Luer Lock
Adapter
Waste
Waste
Superloop
SUPERLOOPS
10 ml
50 ml
Buffer
150 ml
Plunger
BUFFER
FLUSH
SAMPLE
LOAD
SAMPLE
INJECT
SAMPLE
FLUSH
WASTE
PUMP or
SYRINGE
COLUMN
COLUMN
Sample
Etched
Groove
Volume
Max. allowed column
pressure
Code No.
10 ml
50 ml
150 ml
4 Mpa
4 Mpa
2 Mpa
18-1113-81
18-1113-82
18-1023-85
INV-907 Injection Valve - Injection Port
PEEK Valve Body
Injection Needle Port
Valve Seal
Ferrule
NeedleTube
Needle
Needle
Adapter
INV-907 Injection Valve - Luer Adapter
PEEK
PEEKInjection
InjectionValve
ValveBody
Body
PEEK
PEEKInjection
InjectionLuer
LuerAdaptor
Adaptor
Valve
ValveSeal
Seal
Valve
ValveChannel
ChannelPlate
Plate
Fluid Dynamics
Flow
PEEK Tubing
High Resistance
to Flow
Low Resistance
to Flow
End
Beginning
General
Fluid Dynamics
Flow - Injecting a Loop
The way we imagine it Flows
The way it Actually Flows
Bubble Segmentation
100ul Loop
Waste
50ul Load
60ul Load
100ul Load
200ul Load
• Partial Loading Loop = No more than 1/2 Loop Volume
• Total Loading Loop = Inject 2X Loop Volume
LOADING A SAMPLE LOOP
Injection valve
INV-907
1
Column
7
2
Load
3
Inject
Sample
4
Sample
pump
P-950
Pump
6
5
Waste
LOADING A SUPER LOOP
Injection valve
INV-907
1
Column
7
2
Load
Inject
3
Pump
6
4
5
Buffer
Sample
Waste
Sample
MULTI-LOADING A SAMPLE LOOP
Injection valve
INV-907
1
Column
7
2
Load
3
Inject
4
Pump
6
5
PV-908
1
8
2
3
I/O
4
5
7
6
Wash
Samples
Waste
MULTI-LOADING A SUPER LOOP
Column
1
2
Pump
Load
Inject
Injection valve
INV-907
7
3
4
Waste
6
5
PV-908
1
8
2
3
I/O
4
5
7
6
Samples
Wash
S3 S4 S5 S6
S8
DIRECT LOADING ON TO COLUMN
Column
1
Pump
7
2
3
Inject
6
4
Load
5
Extra Tube
1/16” or 1/8”
Tubing
Waste
Sample
REVERSE FLOW VALVE ( INV-907 )
1
7
2
Pump
3
5
4
Column
Extra Tube
6
Stop
Plug
MONITOR
Downward Flow
Position 1
1
7
2
Pump
3
4
Column
Extra Tube
5
6
Stop
Plug
MONITOR
Upward Flow
Position 2
OUTLET VALVE PV-908
DETECTOR
To Fraction
Collector
Default
F8
F1
F2
F3
I/O
F4
F5
F7
F6
F7
Waste
F8
F4
F3
F5
F6
0
250
500
mAU
Flowthrough
Fractionation
Inject
A1 A2 A3………………….
Eluate
Fractionation
0%B
100%B
Peak
Fractionation
End
Fraction Collection
Waste
Start
0
50
100
%B
HiTrap Connections
1/16” Female
1/16” Male
18-1112-55
1/16” Fingertight Fitting
1/16” Female
M6 Male
18-1112-57
M6 male / 1/16” Female
Phenyl Sepharose
HiTrap Column
M6 Male
M6 Female
18-1112-58
M6 female / 1/16” male
Flow Cell Inlet
1/16” Male
Multiple HiTrap Arrangment ( Series )
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
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