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Download BioLogic DuoFlow Chromatography System Starter Kit ...· BioLogic DuoFlow™ Chromatography System

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BioLogic DuoFlowChromatography System

Starter Kit Instruction Manual

Catalog # 760-0135

B L D F

F3 F4 F5

1 2 3

4 5 6

7 8 9

0F1 F2

A B

VALVE A VALVE B

BioFrac Franction Collector

4006208B.qxd 6/28/2004 11:03 AM Page 2

Table of ContentsIntroduction.......................................................................................11. Starter Kit Components ..............................................................12. Materials You Will Need...............................................................1

I. BioLogic DuoFlow System ....................................................3Section 1. DuoFlow System Preparation .........................................31.1 Prime the Workstation Pumps.....................................................51.2 Move the AVR7-3 Inject Valve to the Purge Position....................51.3 Purge the Workstation Pumps ....................................................51.4 Manual Control of the Workstation Pumps ..................................61.5 Flush the System Through to the Fraction Collector ....................6

BioFrac Fraction Collector1.6 Turn on the UV Lamp..................................................................6

DuoFlow UV DetectorQuadTec UV/Vis Detector

1.7 Manual Screen Chromatogram Window......................................71.8 Status Bar...................................................................................7

Section 2. Anion Exchange Separation of Protein Standards........8 2.1 Overview of the Procedure..........................................................82.2 Prepare Buffers...........................................................................92.3 Prepare Sample..........................................................................92.4 Install UNO Q1 Column ............................................................102.5 Prime the Pumps and Equilibrate the UNO Q1 Column ............102.6 Create a New Method...............................................................10

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II. DuoFlow Maximizer and Pathfinder Systems.....................18 Section 3. System Preparation.......................................................183.1 Prime the Workstation Pumps ..................................................203.2 Move the AVR7-3 Inject Valve to the Purge Position..................203.3 Purge the Workstation Pumps ..................................................213.4 Manual Control of the Pumps ...................................................213.5 Flush the System Through to the Fraction Collector ..................21

BioFrac Fraction Collection3.6 Turn on the UV Lamp................................................................22

DuoFlow UV DetectorQuadTec UV/Vis Detector

3.7 pH Electrode Calibration ...........................................................233.8 Manual Screen Chromatogram Window ...................................233.9 Status Bar ................................................................................24

Section 4. Anion Exchange Separation of Protein Standards ......244.1 Overview of the Procedure........................................................244.2 Prepare Solutions .....................................................................254.3 Prepare Sample........................................................................264.4 Install the UNO Q1 Column.......................................................264.5 Prime the Pumps and Equilibrate the UNO Q1 Column.............274.6 Create a New Method...............................................................29

Section 5. Ordering Information.....................................................36

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1

IntroductionThis instruction manual and starter kit contents may be used for the BioLogicDuoFlow system and the BioLogic DuoFlow Maximizer and Pathfinder

chromatography systems. The use of the starter kit with these systems isdescribed in Sections 1 and 3, respectively.

1. Starter Kit ComponentsThis starter kit contains the following items for running a separation:

50 ml of buffer A, 250 mM Tris-HCI buffer, pH 8.1 (10x concentrate)

50 ml of buffer B, 250 mM Tris-HCI buffer, pH 8.1, plus 5.0 M NaCl (10x concentrate)

50 ml of Maximizer solution A1, 500 mM Tris-HCl (10x concentrate)

50 ml of Maximizer solution A2, 500 mM Tris base (10x concentrate)

50 ml of Maximizer solution B2, 5.0 M NaCl (2.5 x concentrate)

One vial of anion exchange protein standard (catalog #125-0561)

One 1 ml disposable sample injection syringe

One 50 l sample loop

The chromatographic separation for this kit requires approximately 6 minutes.

2. Materials You Will NeedIn order to prepare the starter kit buffer solutions you will need the followingmaterials:

Filtered high-quality water (i.e., HPLC grade water)

One 500 ml graduated cylinder

One 1 L side-arm flask

Stirbar and stirplate

Vacuum source for degassing

Two 500 ml bottles

Fraction collection tubes, 13 x 100 mm (at least 14 tubes)

100 ml beaker

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2

If you are using the DuoFlow Maximizer or Pathfinder systems you will also needthe following materials:

pH 7.00 and pH 10.00 standard buffer

One 200 ml graduated cylinder (optional)

Two additional 500 ml bottles

Fraction collection 1.5 or 2 ml micro tubes

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3

I. BioLogic DuoFlow SystemSection 1. DuoFlow System Preparation

When the DuoFlow system is turned on, the Manual screen is displayed (seeFigures 1 and 2). This screen displays instrument control panels that providedirect control of the pumps, valves, fraction collector, UV detector, QuadTec

UV/Vis detector, and Econo Gradient Pump. The arrow button in the upperright corner of the detector control panel toggles between the UV and QuadTecdetector control panels. Only those instruments connected to the system will bedisplayed.

BioLogic Duo-Flow - - - no method -

Edit

MethodNew

MethodNew

Run Browser Manual Setup SetupProtocolwashload

12

Run NotesReport PostRun Log

Advance

Fraction Collector: BioFrac

START

Gradient Pump: F10

Inlet A

1.00

50

50Inlet B

Flowrate ml/min

%

%

Highlimit

0

700

Lowlimit psi.

AVR7-3 at port 4

Workstation Valves

SVT5-4 at port 1 SVT3-2 at port 3SV5-4 at port 2

PL21

21

34

21

34

STOP START STOPSTART STOP

Set

Event Mark

OFFON

psi.

Econo Gradient Pump 1

EGP %B

1.000

0

0.00% Split

FlowDirection

Flowrate ml/min

%

%

Set

Tube number:

ml

Volume left:

Mode: System Local

Conductivity range(mS/cm):

Mode: System Local

Start Tube:

End Tube:

Fraction size:

1

1

20

1.00

5

1.0

I

AVR9-8 at port 5

37

8 2

65

4

I

AVR9-8 at port 6

37

8 2

65

4

I

UV Detector

Chart Recorder

ON OFF

Zero Baseline

100% Buffer B

0 2 4 6 8 10MinutesAU mS/cm

1.50

1.00

0.50

50

0.00 0.0

400.0

300.0

200.0

100.0

FractionsUV Conductivity

ClearTraces

Resize

UV range (AU):

Flow Rate EGP %B

0 %B0.00 ml/min 0%

% Split

0.00 ml/min 50 %B

Gradient Pump: F10 UV Conductivity

1 psi 0.2583 AU 0.000 mS/cm

QuadTec Econo GradientPump

SIM1/pHSIM1/SIG

Settings

File View Utilities Options Window HelpBio-Rad

Web

Rack: F1 (12-13 mm tubes)

Zero baseline for the UV detector

Fig. 1. Manual Control Screen with UV detector

4006208B.qxd 6/28/2004 11:03 AM Page 8

4

BioLogic Duo-Flow - - - no method -

Edit

MethodNew

MethodNew

Run Browser Manual Setup SetupProtocolwashload

12

Run NotesReport PostRun Log

Advance

Fraction Collector: BioFrac

START

Gradient Pump: F10

Inlet A

1.00

50

50Inlet B

Flowrate ml/min

%

%

Highlimit

0

700

Lowlimit psi.

AVR7-3 at port 4

Workstation Valves

SVT5-4 at port 1 SVT3-2 at port 3SV5-4 at port 2

PL21

21

34

21

34

STOP START STOPSTART STOP

Set

psi.

Econo Gradient Pump 1

EGP %B

1.000

0

0.00% Split

FlowDirection

Flowrate ml/min

%

%

Set

Tube number:

ml

Volume left:

Mode: System Local Mode: System Local

Start Tube:

End Tube:

Fraction size:

1

1

20

1.00

I

AVR9-8 at port 5

37

8 2

65

4

I

AVR9-8 at port 6

37

8 2

65

4

I

ON OFF

Zero Baseline

100% Buffer B

0 2 4 6 8 10MinutesAU mS/cm

1.50

1.00

0.50

50

0.00 0.0

400.0

300.0

20

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