BIO RAD BioLogic Duo-Flow Starter Kit Instructions
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BIO RAD BioLogic Duo-Flow Starter Kit Instructions

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BioLogic
Chromatography System
Starter Kit Instructions
for the First Time User of the
BioLogic Duo-Flow and
BioLogic HR Systems
Catalog Number
750-0135
For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)

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  • Page 1 BioLogic Chromatography System Starter Kit Instructions for the First Time User of the BioLogic Duo-Flow and BioLogic HR Systems Catalog Number 750-0135 For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)

  • Page 2: Table Of Contents

    Introduction ...............1 Starter Kit Components ..............1 Other Materials You Will Need ............2 Part 1 BioLogic Duo-Flow Chromatography System Section 1 Preparation for Use of the System ........3 Section 2 Anion Exchange Separation of Protein Standards ..7 Overview of the Procedure ............7 Buffer Preparation ................8...

  • Page 3: Introduction

    This instruction manual and starter kit contents may be used for both the BioLogic Duo-Flow and BioLogic HR Chromatography Systems. To use the starter kit with the BioLogic Duo-Flow System, refer to Section 1. To use the starter kit with the BioLogic HR System, refer to Section 3.

  • Page 4: Other Materials You Will Need

    2. Other Materials You Will Need • Filtered high quality water (i.e., HPLC grade water) • One 500 ml graduated cylinder • One 1 L sidearm flask • Stir bar and stir plate • Vacuum source for degassing • Two 500 ml bottles •...
  • Page 5 Preparation for Use of the Duo-Flow System When the BioLogic Duo-Flow System is turned on, the Manual Control Screen is displayed. (see Figure 1) This screen consists of a series of boxes containing instrument faceplates which provide direct control of the gradient pumps, a fraction collector, the UV and conductivity detector range settings and valves.
  • Page 6 The gradient pumps will run and the indicator lights will flash green. c. Run both pumps for 2 minutes and then press the purge buttons again to stop the pump. BioLogic Duo-Flow - Default - no method File View Utilities...
  • Page 7 Connect the Model 2128 Fraction Collector by inserting the gray bus cable between it and the workstation. When it is connected, it will appear in the Manual Screen, as shown in Figure 2. BioLogic Duo-Flow - Default - no method File View...
  • Page 8 The Model 2128 Fraction Collector has two operating modes. • System: Controlled by the BioLogic Duo-Flow System. • Local: Controlled from its own faceplate. For now, insure that the system button is selected. When in System mode, the fraction collector operates as follows.

  • Page 9: Section 2 Anion Exchange Separation Of Protein Standards

    Section 2 Anion Exchange Separation of Protein Standards The starter kit allows you to learn how to use the BioLogic Duo-Flow System by programming and running a separation of a premixed anion exchange standard containing equine myoglobin, conalbumin, chicken ovalbumin and soybean trypsin inhibitor, using a 1.3 ml UNO Q1 Column Equine myoglobin is not retained on the UNO Q1 Column and elutes in the void volume.

  • Page 10: Buffer Preparation

    2.2 Buffer Preparation Buffer A 1. Empty the contents of the bottle labeled buffer A into a 500 ml graduated cylinder and add filtered, high-quality water to 500 ml volume. 2. Place the contents of the graduated cylinder into a 1 L side-arm flask and drop in a stir bar.

  • Page 11: Priming The Gradient Pumps And Equilibrating The Uno Q1 Column

    The initial default Devices in Setup are a UV detector and a conductivity monitor as these come as standard with the BioLogic Duo-Flow System. The default devices in Setup can be changed by choosing Save setup as default from the...
  • Page 12 You will see that the valve position names are Load for position 1, Inject for position 2, and Purge for position 3. BioLogic Duo-Flow - Jennifer - System training - Starter Kit - File Edit...
  • Page 13 Programming the Protocol Editor 1. From the Options pull-down menu, insure that Use Volume (ml) is selected, so that the programming base is Volume. BioLogic Duo-Flow - Dr. J. Smith - Starter Kit - UNO Q1 - Edit View Utilities...
  • Page 14 Step Start Number (ml) Step Collect fractions of size 2.00 ml during entire run Isocratic flow with 100% 25 mM tris, pH 8.1, 0% 25 mM tris, 0.5 M NaCl, pH 8.1 at 4.00 ml/min for 1.0 ml Set UV baseline to 0.0 Static loop: Inject 0.5 ml sample at 4.00 ml/min for 0.1 min Isocratic flow with 100% 25 mM tris, pH 8.1,...
  • Page 15 The Run Control Screen 1. Use the toolbar buttons at the left-hand side of the screen to check that the screen display ranges for UV (0.1 AUFS) and conductivity (100 mS/cm) are correctly set and that the gradient pump pressure limits are appropriate (set to 700 psi High and 20 psi Low).
  • Page 16 BioLogic Duo-Flow - Carole Smith - Uno Q1 data - Uno Q1 - Standards Window File View Utilities Options Help wash Edit load Browser Method Method Report Manual Setup Protocol Notes PostRun FullView Conductivity Frac. Collector Fractions Advance 0.100 50.0...
  • Page 17 Section 3 Preparation for Use of the BioLogic HR System The procedures in this document are written for two configurations, which are distinguished by the following. • Manual: Uses a V7-3 Manual Inject Valve, a Model 2110 Fraction Collector, and an SV3-2 Diverter Valve. •...
  • Page 18 If you plugged an inject valve into port 4, you will see a valve box designated AV7-3 at port 4. The three radio buttons of this box correspond to valve positions as follows: L = Load position, I = Inject position, P = Purge position. To move the AV7-3 valve to Purge position, click button P.
  • Page 19 4. Control of the BioLogic HR Pumps. The gradient pump parameters are set from the manual screen either by clicking in the appropriate field and entering a value from the keyboard or by using the spin arrows. You can alter the flow rate from 0.1 to 10 ml/min for any given proportion of pumps A and B.
  • Page 20 • Continue to pump water through the system for at least 10 minutes. You may wish to continue fraction collection or return the SV3-2 valve to position 1 and then stop fraction collection. Note: In manual operation, starting or stopping fraction collection does not change the SV3-2 diverter valve position.
  • Page 21 6. Turning on the UV lamp. a. The UV lamp automatically turns on when you turn on power to the workstation. The UV lamp is turned off and on by clicking on the toggle button. You should now check that the lamp is on; the mercury lamp requires approximately 30 minutes to warm up.
  • Page 22 9. Controlling the chart recorder. Insure that the voltage inputs on the recorder are set at 1V. • The chart recorder pen up/down and chart stop/start functions are controlled by the ON/OFF toggle switch, which is displayed in the Manual screen. •...

  • Page 23: Section 4 Anion Exchange Separation Of Protein Standards

    Section 4 Anion Exchange Separation of Protein Standards The starter kit allows you to learn how to use the BioLogic HR System by programming and running a separation of a premixed anion exchange standard containing equine myoglobin, conalbumin, chicken ovalbumin and soybean trypsin inhibitor, using a 1.3 ml UNO Q1 Column .
  • Page 24 4.2 Buffer Preparation Buffer A 1. Empty the contents of the bottle labeled buffer A into a 500 ml graduated cylinder and add filtered, high-quality water to 500 ml volume. 2. Place the contents of the graduated cylinder into a 1 L side-arm flask and drop in a stir bar.

  • Page 25: Priming The Gradient Pumps And Equilibrating The Uno Q1 Column

    4.5 Priming the Gradient Pumps and Equilibrating the UNO Q1 Column Be sure the gradient pumps are stopped and the inject valve is in the purge position. Re-prime and purge pumps A and B as described in Section 2, step 1 of this manual. Set the inject valve to position L (Load).
  • Page 26 Programming the Setup Editor The information entered in the setup editor screen is important for the subsequent separation method. The buttons grouped in the left hand box (Available Devices) show all the devices that could theoretically be connected to the BioLogic HR System. The list of items in the right hand box (Devices in Setup) identifies those devices you wish to use in the subsequent method.
  • Page 27 1. Click on the fraction collector button in the Available Devices box. A dialog box will appear asking you to choose the type of collector (a generic collector, a Model 2110 or a Model 2128). Click on Model 2110 and click the OK button. You will now see 2110 Fraction Collector in the Devices in Setup box (see Figure 3).
  • Page 28 2. Program the separation method presented in the table on the following page. • From the left side of the screen, click on the Fraction Collection button. In the pop-up window that appears, choose Collect All with a fraction size of 2.00 ml and a delay of 0. •...
  • Page 29 Step (ml) Number Start Step Collect fractions of size 2.00 ml during entire run Turn chart recorder ON Isocratic Flow with 100% 25 mM tris, pH 8.1, 0% 25 mM tris, 0.5 M NaCl, pH 8.1 at 4.00 ml/min for 1.0 ml Set UV baseline to 0.0 Static loop: Inject 0.5 ml sample at 4.00 ml/min for 0.1 min...
  • Page 30 The Run Control Screen 1. Use the toolbar buttons at the left side of the screen to check that the chart recorder ranges for UV (0.1 AUFS) and conductivity (100 mS/cm) are correctly set and that the gradient pump pressure limits are appropriate (set to 700 psi High and 20 psi Low).
  • Page 31 Starting the Run 1. Insure that sufficient tubes are in the fraction collector rack (approximately 14) and that the drop-head is over tube 1. 2. Insure that the V7-3 valve is in the LOAD position (position L). 3. Insure that the 50 µl sample loop is connected to ports 3 and 6 of the inject valve.

  • Page 32: Creating A Method For The Automated Biologic Hr System

    4.7 Creating a Method for the Automated BioLogic HR System From the Manual Control Screen, click the New Method toolbar button. You will see a dialog box asking for a method name (input Starter Kit Separation–Auto). Use the keyboard tab key to go to the Method Description and Method Author fields.
  • Page 33 BioLogicHR - Jennifer - System training - Starter Kit - File Edit View Utilities Options Window Help wash Edit load Browser Method Method Report Manual Setup Protocol Notes PostRun Delete Devices in setup Available Devices 2128 Fraction Collector Aux Load Fraction UV Detector Pump...
  • Page 34 Programming the Protocol Editor 1. From the Options pull-down menu, insure that Use Volume (ml) is selected, so that the programming base is Volume. BioLogicHR - Dr. J. Smith - Starter Kit - UNO Q1 - Edit View Utilities Options Window Help File...
  • Page 35 Step Start Number (ml) Step Collect fractions of size 2.00 ml during entire run Turn chart recorder ON Isocratic flow with 100% 25 mM tris, pH 8.1, 0% 25 mM tris, 0.5 M NaCl, pH 8.1 at 4.00 ml/min for 1.0 ml Set UV baseline to 0.0 Static loop: Inject 0.5 ml sample at 4.00 ml/min for 0.1 min...
  • Page 36 The Run Control Screen 1. Use the toolbar buttons at the left side of the screen to check that the chart recorder ranges for UV (0.1 AUFS) and conductivity (100 mS/cm) are correctly set and that the gradient pump pressure limits are appropriate (set to 700 psi High and 20 psi Low).
  • Page 37 BioLogicHR - Carole Smith - Uno Q1 data - Uno Q1 - Standards File View Utilities Options Window Help wash Edit load Browser Method Method Report Manual Setup Protocol Notes FullView PostRun Conductivity Frac. Collector Fractions Advance 0.100 50.0 100% Buffer B Divert Valve 0.090 45.0...
  • Page 38 Bio-Rad Laboratories Life Science Website www.bio-rad.com U.S. (800) 4BIORAD Australia 02 9914 2800 Austria (01) 877 89 01 Belgium 09-385 55 11 Canada (905) 712-2771 Group China 86-10-62051850 86-10) 62051850 Denmark 45 39 17 99 47 Finland 358 (0)9 804 2200 France (01) 43 90 46 90 Germany 089 318 84-0 Hong Kong 852-2789-3300 India (91-11) 461 0103 Israel 03 951 4127 Italy 02 21609.1 Japan 03-5811-6270 Korea 82-2-3473-4460...

This manual is also suitable for:

Biologic hr750-0135

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