Bio-rad ChromLab™ Software Bedienungsanleitung

NGC™ Chromatography Systems and ChromLab™ SoftwareUser GuideVersion 3.03.0

viii | NGC Chromatography Systems and ChromLab SoftwareTable of ContentsAnnotating the Chromatogram . . . . . . . . . . . . . . . . . . . . . .

4 | Performing a Manual Run98 | NGC Chromatography Systems and ChromLab Software3. To purge an individual line of air bubbles and fill it

Preparing the SystemUser Guide | 9 9To purge the buffer inlet valves (if available)1. Open the System Pump dialog box and select Priming on the Mo

4 | Performing a Manual Run100 | NGC Chromatography Systems and ChromLab SoftwareTo purge the sample pump (if available)1. In the fluidic

Preparing the SystemUser Guide | 101To purge sample inlet valves (if available)1. In the fluidic scheme, double-click the Sample Pump module to op

4 | Performing a Manual Run102 | NGC Chromatography Systems and ChromLab SoftwareCleaning the SystemSystem cleaning consists of rinsing th

Running an ExperimentUser Guide | 103Running an ExperimentImportant: When running an experiment manually, ensure that the valves are set in the c

4 | Performing a Manual Run104 | NGC Chromatography Systems and ChromLab SoftwareTo set up the experiment in the fluidic scheme1. Double-c

Running an ExperimentUser Guide | 1054. (Optional) Double-click the Column Switching Valve module, if installed, choose the following settings, an

4 | Performing a Manual Run106 | NGC Chromatography Systems and ChromLab Software6. Double-click the Sample Pump module, if installed, and

Running an ExperimentUser Guide | 1077. Double-click the System Pump module and choose the following settings in the dialog box that appears:a. If

User Guide | i xTable of ContentsPeak Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229Peak

4 | Performing a Manual Run108 | NGC Chromatography Systems and ChromLab SoftwareTo run an experiment step1. Isocratic or gradient step:a.

Running an ExperimentUser Guide | 109Changing Module SettingsTip: For some modules, you must stop the pumps before you can change the settings.T

4 | Performing a Manual Run110 | NGC Chromatography Systems and ChromLab SoftwareSaving a Manual Run on the Touch ScreenTo save a manual r

Running an ExperimentUser Guide | 111Viewing Run DataNote: See Chapter 7, Evaluating Results, for detailed information about analyzing a run.When

4 | Performing a Manual Run112 | NGC Chromatography Systems and ChromLab Software

User Guide | 1135 Method EditorThis chapter introduces ChromLab™ software Method Editor concepts and functionality and prepares you to create a me

5 | Method Editor114 | NGC Chromatography Systems and ChromLab SoftwareBefore You BeginBefore you work with Method Editor features, it is im

Accessing Method Editor FeaturesUser Guide | 115Opening a MethodWhen you click Open Method in the Method Editor pane, the Open Method dialog box a

5 | Method Editor116 | NGC Chromatography Systems and ChromLab SoftwareMethod Editor WindowThe Method Editor enables you to create, open, im

Method Editor WindowUser Guide | 117File Menu CommandsNew Method — opens the Method Editor window in which you can create a method. Open Method —

x | NGC Chromatography Systems and ChromLab SoftwareTable of ContentsChapter 8 Importing and Exporting Data . . . . . . . . . . . . . . . . .

5 | Method Editor118 | NGC Chromatography Systems and ChromLab SoftwareExport Method — exports a saved regular or scouting method to the des

Method Editor WindowUser Guide | 119View Menu CommandsPhase Library – displays the Phase Library pane. This is the default. Clearing this command

5 | Method Editor120 | NGC Chromatography Systems and ChromLab SoftwarePhase Library PaneThe Phase Library pane contains both standard and c

Phase Library PaneUser Guide | 121Adding a Phase to a MethodTo add a phase to a method Do one of the following: Drag the phase to the appropriat

5 | Method Editor122 | NGC Chromatography Systems and ChromLab SoftwareMethod Outline PaneThe Method Outline pane lists phases in the order

Method Steps PaneUser Guide | 123In this pane you can define the parameters of each method phase. Parameters vary depending on the phase and the d

5 | Method Editor124 | NGC Chromatography Systems and ChromLab SoftwareViewing Details of a Step in the Step TableTo view step details Righ

Method Settings ParametersUser Guide | 125Method Settings ParametersMethod Settings are general parameters that affect the entire method. These se

5 | Method Editor126 | NGC Chromatography Systems and ChromLab SoftwareColumn SelectionUnder Column Selection you select the column and the

Method Settings ParametersUser Guide | 127You can remove a column from the list if it is no longer used, clearing the assigned port. You can then

User Guide | 1 11 IntroductionNGC™ chromatography systems provide a general purpose purification platform for purifying all forms of biomolecules

5 | Method Editor128 | NGC Chromatography Systems and ChromLab Software3. Select a column position to configure from the dropdown list.4. (O

Method Settings ParametersUser Guide | 129To add user-defined columns1. In the Column Selection section, do one of the following: Select Single C

5 | Method Editor130 | NGC Chromatography Systems and ChromLab Software2. Provide the requisite information for the column. A red asterisk (

Method Settings ParametersUser Guide | 131To remove user-defined columns1. In the Column Selection section, do one of the following: Select Singl

5 | Method Editor132 | NGC Chromatography Systems and ChromLab SoftwarepH Valve Enable pH monitoring — if a pH valve is present, this checkb

Method Settings ParametersUser Guide | 133BioFrac Settings — by default, fraction collection begins in the first tube of the first rack. The fract

5 | Method Editor134 | NGC Chromatography Systems and ChromLab Software Buffer Blending — buffer acid, buffer base, water, and salt are mix

Method Settings ParametersUser Guide | 135Select Buffer — opens the Buffer Recipes dialog box in which you can choose a recipe for the buffer and

5 | Method Editor136 | NGC Chromatography Systems and ChromLab SoftwareAvailable Range — available ranges for the recipe concentration and p

Phase Controls and ParametersUser Guide | 137In gradient segments, lower numbers yield increasing gradients. Higher numbers yield negative gradien

1 | Introduction12 | NGC Chromatography Systems and ChromLab SoftwareChromLab™ software enables you to set up and control an NGC instrument,

5 | Method Editor138 | NGC Chromatography Systems and ChromLab SoftwareStandard PhasesSome standard phases use identical parameters. For exa

Phase Controls and ParametersUser Guide | 139Loading the Loop with a Sample PumpWhen a sample pump is used to load the loop, you can define the fl

5 | Method Editor140 | NGC Chromatography Systems and ChromLab SoftwareYou can change the port on the sample inlet valve from which to load

Phase Controls and ParametersUser Guide | 141Injecting Sample Directly with a Sample PumpWhen a sample pump is present in the fluidic scheme and t

5 | Method Editor142 | NGC Chromatography Systems and ChromLab SoftwareYou can change the port on the sample inlet valve from which to load

Phase Controls and ParametersUser Guide | 143Injecting Sample with an AutosamplerIf your fluidic scheme includes an autosampler and you choose to

5 | Method Editor144 | NGC Chromatography Systems and ChromLab SoftwareAlthough it is typically used after the sample application or elution

Phase Controls and ParametersUser Guide | 145System CIP (Clean in Place) — Storage ParametersNote: System CIP must be run as the only type of pha

5 | Method Editor146 | NGC Chromatography Systems and ChromLab Software NGC Scout SystemsBuffer selection: Manually Prepared Buffer via Blen

Phase Controls and ParametersUser Guide | 147NGC Discover and NGC Discover Pro SystemsNote: To perform CIP for the buffer blending valve and buff

Main NGC FeaturesUser Guide | 1 3 Organize the location of the modules to optimize separation performance based on method scale and complexity, a

5 | Method Editor148 | NGC Chromatography Systems and ChromLab Software Sample inlet valves are washed by using the sample pump to pull was

Phase Controls and ParametersUser Guide | 149 Wash solution is delivered through buffer inlet ports for the sample loop, pH flow cell, and column

5 | Method Editor150 | NGC Chromatography Systems and ChromLab Software Column switching valve Outlet valve Sample inlet valves Wash sol

Phase Controls and ParametersUser Guide | 1516. Fill a syringe with distilled water, connect the syringe to the pH valve Cal In port, and inject t

5 | Method Editor152 | NGC Chromatography Systems and ChromLab SoftwareColumn Preparation/Activation ParametersThis phase is used to prepare

Phase Controls and ParametersUser Guide | 153Column Performance Test ParametersThis test combines the three basic phases necessary for a run (equi

5 | Method Editor154 | NGC Chromatography Systems and ChromLab SoftwareSystem Preparation ParametersThis phase removes storage solution and

Phase Controls and ParametersUser Guide | 155Create New Phase ParametersThis phase enables you to create a new phase by adding steps from the Step

5 | Method Editor156 | NGC Chromatography Systems and ChromLab SoftwareStep Library PaneVisible when the Method Steps tab is selected, the S

Scout Parameters PaneUser Guide | 157 Zero Baseline Hold Until Pause System WashTo get more information about method steps In the Method Edit

1 | Introduction14 | NGC Chromatography Systems and ChromLab SoftwareNGC Chromatography SystemsAll NGC chromatography systems include ChromL

5 | Method Editor158 | NGC Chromatography Systems and ChromLab Software

User Guide | 1596 Creating a MethodIn the NGC™ chromatography system, methods are used to encapsulate an entire process to be run. Methods consist

6 | Creating a Method160 | NGC Chromatography Systems and ChromLab SoftwareStandard Method TemplatesThe workflow for standard method templat

Standard Method TemplatesUser Guide | 161Chromatofocusing Chromatofocusing chromatography uses a charged matrix to bind protein molecules. A pH g

6 | Creating a Method162 | NGC Chromatography Systems and ChromLab SoftwareMulticolumn Tandem Multicolumn tandem purification uses these tem

Creating a Method from a TemplateUser Guide | 163Creating a Method from a Template Standard methods are supplied as templates. You must save a tem

6 | Creating a Method164 | NGC Chromatography Systems and ChromLab Software2. In the left pane, click a technique folder and select a templa

Creating a Method from Standard PhasesUser Guide | 1656. Do one of the following: Select a project folder. In the Method Name box, type a name fo

6 | Creating a Method166 | NGC Chromatography Systems and ChromLab SoftwareEach element of the fluidic scheme is associated with settings in

Creating a Method from Standard PhasesUser Guide | 1672. Choose a fluidic scheme that matches your NGC instrument configuration and click Select.

NGC Chromatography SystemsUser Guide | 1 5The NGC Scout™ chromatography system includes the following: All modules on the Quest system pH detect

6 | Creating a Method168 | NGC Chromatography Systems and ChromLab Software5. Depending on the method, either Single Wave UV Detector Settin

Creating a Method from Standard PhasesUser Guide | 169Serpentine is selected by default. Fractions can also be collected by rows or columns for mi

6 | Creating a Method170 | NGC Chromatography Systems and ChromLab Softwareabout priming your system.9. Select a pump head type. To set the

Renaming PhasesUser Guide | 1713. Repeat steps 1 and 2 until you have finished editing phase settings for the method.4. Save the method to preserv

6 | Creating a Method172 | NGC Chromatography Systems and ChromLab SoftwareDeleting PhasesNote: You cannot delete the Method Settings phase

Running a Method and Collecting FractionsUser Guide | 173Running a Method and Collecting FractionsYou can schedule a method to start immediately o

6 | Creating a Method174 | NGC Chromatography Systems and ChromLab SoftwareCollecting Fractions in Run QueuesWhen you schedule queued method

Running a Method and Collecting FractionsUser Guide | 175To run a method1. Open the method to run.2. On the toolbar, click Start Run. The Schedule

6 | Creating a Method176 | NGC Chromatography Systems and ChromLab Software3. Repeat steps 1–2 to add as many methods to the queue as needed

Saving a MethodUser Guide | 177Saving a MethodYou can modify a method — whether or not you have run it — and save it under the same name. The modi

1 | Introduction16 | NGC Chromatography Systems and ChromLab SoftwareFinding Out MoreAfter you install NGC documentation from the NGC Chroma

6 | Creating a Method178 | NGC Chromatography Systems and ChromLab Software2. Type a name for the new template and click Save.Renaming a Met

Deleting a MethodUser Guide | 179Deleting a MethodYou can delete an open method. If the method is associated with runs, you can choose to delete o

6 | Creating a Method180 | NGC Chromatography Systems and ChromLab SoftwareScouting on Parameters within a MethodScouting is a procedure use

Scouting on Parameters within a MethodUser Guide | 181The Scouting WizardThe scouting wizard guides you through three pages: Choose a Scouting Par

6 | Creating a Method182 | NGC Chromatography Systems and ChromLab Software Column — test up to five column types for each column-switching

Scouting on Parameters within a MethodUser Guide | 183Depending on the parameter you chose on Page 1, you can select individual steps to scout or

6 | Creating a Method184 | NGC Chromatography Systems and ChromLab Softwarecheckbox is also cleared. The other steps remain selected and wil

Scouting on Parameters within a MethodUser Guide | 185 Run Name — the scout run name. The autogenerated name is based on the parameter you chose

6 | Creating a Method186 | NGC Chromatography Systems and ChromLab SoftwareOn this page you can select up to five columns for each column-sw

Scouting on Parameters within a MethodUser Guide | 187 Port — the column’s position on the column switching valve module. This field is not edita

User Guide | 1 72 The WorkspaceChromLab™ software provides an intuitive interface for developing chromatography methods, operating an NGC™ instrum

6 | Creating a Method188 | NGC Chromatography Systems and ChromLab Softwareinclude a preinjection sample pump wash, the method is updated an

Scouting on Parameters within a MethodUser Guide | 189 Include in Scout Sequence — determines whether the run is included in the scout method. By

6 | Creating a Method190 | NGC Chromatography Systems and ChromLab SoftwareSpecial ConsiderationsRules for generating scout sequences can va

Scouting on Parameters within a MethodUser Guide | 191Generating Column Scout Sequences This option is available only when the fluidic scheme inc

6 | Creating a Method192 | NGC Chromatography Systems and ChromLab SoftwareViewing a Summary of the Scouting MethodWhen you complete the sco

Scouting on Parameters within a MethodUser Guide | 193When you collect fractions during scouting, the fractions get appended to the fractions coll

6 | Creating a Method194 | NGC Chromatography Systems and ChromLab Software4. Do one of the following: Click Start Run to begin the run imm

User Guide | 1957 Evaluating ResultsIn the Evaluation window, you can process and analyze chromatography data. The results can be viewed in a vari

7 | Evaluating Results196 | NGC Chromatography Systems and ChromLab SoftwareManaging Analysis ProjectsYou can open a run or analysis in the

Managing Analysis ProjectsUser Guide | 197To group the runs list by method Right-click the runs list and choose Group by Method in the menu that

2 | The Workspace18 | NGC Chromatography Systems and ChromLab SoftwareThe Home WindowChromLab software opens with the Home window, which dis

7 | Evaluating Results198 | NGC Chromatography Systems and ChromLab SoftwareEvaluation WindowIn the Evaluation window you can display run da

Evaluation WindowUser Guide | 199File Menu CommandsOpen Run/Analysis — opens the Open Run/Analysis dialog box in which you can select runs to disp

7 | Evaluating Results200 | NGC Chromatography Systems and ChromLab SoftwareImport Unicorn Data — opens a dialog box in which you can import

Evaluation WindowUser Guide | 201View Menu CommandsOverlay View — displays multiple runs in a single chromatogram window.Stack View — displays mul

7 | Evaluating Results202 | NGC Chromatography Systems and ChromLab SoftwareDelete Peak List — deletes previously calculated peaks. All peak

Customizing the ChromatogramUser Guide | 203Customizing the Chromatogram You can change the color of the traces, show or hide selected traces, con

7 | Evaluating Results204 | NGC Chromatography Systems and ChromLab SoftwareSelecting the Active TraceWhen a trace is selected, it appears b

Customizing the ChromatogramUser Guide | 205Showing or Hiding a TraceTo show or hide a trace In the trace table, select or clear the Show checkbo

7 | Evaluating Results206 | NGC Chromatography Systems and ChromLab SoftwareTo rescale UV units individually for multiple or overlaid traces

Customizing the ChromatogramUser Guide | 207To revert to the default trace color Right-click the trace color in the table and choose Restore Defa

The Home WindowUser Guide | 1 9Method Editor and Evaluation tabs become visible in the Home window when you select tasks in the Method Editor and

7 | Evaluating Results208 | NGC Chromatography Systems and ChromLab SoftwareObtaining Statistics of a Selected Region on the ChromatogramFor

Customizing the ChromatogramUser Guide | 209To select a region on the chromatogram Right-click the start point in the chromatogram and drag the p

7 | Evaluating Results210 | NGC Chromatography Systems and ChromLab SoftwareTo clear the selected region on the chromatogram Close the Regi

Customizing the ChromatogramUser Guide | 211To edit an annotation1. Double-click the annotation on the chromatogram to open its dialog box.2. Edit

7 | Evaluating Results212 | NGC Chromatography Systems and ChromLab SoftwareCustomizing the Trace Table DisplayThe details of each trace in

Customizing the Trace Table DisplayUser Guide | 213Sorting Table ColumnsWhen the trace table is not grouped, traces are sorted in default order. T

7 | Evaluating Results214 | NGC Chromatography Systems and ChromLab SoftwareCopying a TableYou can copy a table to the clipboard and then pa

Customizing the Trace Table DisplayUser Guide | 215Manually Setting the Injection Point (x Alignment)You can manually set the injection point for

7 | Evaluating Results216 | NGC Chromatography Systems and ChromLab SoftwareManaging RunsYou can rename or delete a single run displayed in

Managing RunsUser Guide | 217Viewing Multiple Runs in Separate TabsChromLab software can display multiple runs in the Evaluation window. Each run

2 | The Workspace20 | NGC Chromatography Systems and ChromLab SoftwareFile Menu CommandsConnect to System — opens a dialog box that enables

7 | Evaluating Results218 | NGC Chromatography Systems and ChromLab SoftwareComparing TracesChromLab can display multiple runs selected in t

Comparing TracesUser Guide | 219Viewing Multiple Runs in One TabIn this view, a single table displays information for all traces in all the displa

7 | Evaluating Results220 | NGC Chromatography Systems and ChromLab Software

Comparing TracesUser Guide | 221Overlay ViewAll visible traces from all selected runs appear in a single chromatogram.To switch between stacked an

7 | Evaluating Results222 | NGC Chromatography Systems and ChromLab SoftwareCustomizing the Trace Comparison TableMultiple runs in a single

Comparing TracesUser Guide | 223Offsetting Traces — Overlay ModeWhen two or more traces from different runs are too close together to distinguish,

7 | Evaluating Results224 | NGC Chromatography Systems and ChromLab SoftwareManaging AnalysesRemoving Runs from a Trace ComparisonWhen you v

Managing AnalysesUser Guide | 225To save an analysis1. Do one of the following: Click Save Analysis on the Run tab toolbar.  Select File > Sa

7 | Evaluating Results226 | NGC Chromatography Systems and ChromLab SoftwareDeleting an AnalysisTo delete an analysis1. With the analysis di

Peak IntegrationUser Guide | 227Peak IntegrationChromLab software uses an algorithm to find peaks in UV traces and calculate the necessary data fo

The Home WindowUser Guide | 2 1Import — displays links from which you can import the following: NGC File — opens a dialog box in which you can im

7 | Evaluating Results228 | NGC Chromatography Systems and ChromLab SoftwareAutomatic Peak Integration SettingsIn the Peak Integration pane,

Peak IntegrationUser Guide | 229Peak ParametersYou can specify initial values for two parameters that participate in the peak detection process: s

7 | Evaluating Results230 | NGC Chromatography Systems and ChromLab SoftwareIntegrate and DeleteIntegrate — detects peaks, using current par

Peak IntegrationUser Guide | 231Displaying Peak Results — the ChromatogramIn the chromatogram, a peak’s start, end, and apex are indicated by smal

7 | Evaluating Results232 | NGC Chromatography Systems and ChromLab SoftwareThe calculated baseline appears on the chromatogram when peaks a

Peak IntegrationUser Guide | 233To hide or show peak view options Right-click in the chromatogram, choose View, and select the option or options

7 | Evaluating Results234 | NGC Chromatography Systems and ChromLab Software Peak asymmetry — the asymmetry factor, defined as the distance

Peak IntegrationUser Guide | 235 Molecular mass (kDa) — the molecular mass of the protein. By default this field is empty. Molecular mass values

7 | Evaluating Results236 | NGC Chromatography Systems and ChromLab SoftwareTo display a hidden column1. Right-click a column heading and se

Peak IntegrationUser Guide | 237To expand or collapse groups of peaks Right-click the table and select Expand All Groups or Collapse All Groups o

2 | The Workspace22 | NGC Chromatography Systems and ChromLab SoftwareHelp Menu CommandsHelp — displays screen-level help topics and links t

7 | Evaluating Results238 | NGC Chromatography Systems and ChromLab SoftwareAdjusting Peaks Manually You can manually adjust the peaks found

Peak IntegrationUser Guide | 239You can adjust peaks by selecting commands on the menu that appears when you right-click in the chromatogram or Pe

7 | Evaluating Results240 | NGC Chromatography Systems and ChromLab SoftwareTo split a peak In the chromatogram or in the Peaks table, righ

Evaluating FractionsUser Guide | 241Evaluating FractionsWhen fraction collection is enabled the details of each fraction in the run appear in a ro

7 | Evaluating Results242 | NGC Chromatography Systems and ChromLab Software Orange — for fractions collected via an outlet valve. The outl

Evaluating FractionsUser Guide | 243Displaying Fraction Collection Results — the Fractions TableLike the Traces table, the Fractions table is avai

7 | Evaluating Results244 | NGC Chromatography Systems and ChromLab Software Extinction coefficient ((mg/ml)-1 cm-1) — the extinction coeff

Evaluating FractionsUser Guide | 245Calculating Protein Concentration for FractionsYou can calculate and view the protein concentration for indivi

7 | Evaluating Results246 | NGC Chromatography Systems and ChromLab SoftwareViewing Fraction DetailsTo view details of a single fraction Do

Evaluating FractionsUser Guide | 247 Tube location — the location of the group of pooled tubes in the rack. Start —the beginning of the collecti

The System Control WindowUser Guide | 2 3The System Control WindowThe System Control window enables you to run the instrument manually, monitor me

7 | Evaluating Results248 | NGC Chromatography Systems and ChromLab SoftwareViewing Fraction Collection Results — the Heat MapA configuratio

Evaluating FractionsUser Guide | 249The color scale is based on the column chosen and is relative among all available fractions of the run. A dark

7 | Evaluating Results250 | NGC Chromatography Systems and ChromLab SoftwareTo hide the rack display Click the collapse arrow to the right

Column Performance AnalysisUser Guide | 251Column Performance AnalysisMethod runs that include the Column Performance Test phase can be analyzed f

7 | Evaluating Results252 | NGC Chromatography Systems and ChromLab SoftwareTracesYou can choose to analyze the column’s performance based o

Column Performance AnalysisUser Guide | 253Reduced HETP (rHETP) — the reduced plate height, a value used to compare the efficiency of multiple col

7 | Evaluating Results254 | NGC Chromatography Systems and ChromLab Software4. In the section, enter the values for bed height and particle

Column Performance AnalysisUser Guide | 255Column Performance — Manual AdjustmentIn the Manual Adjustment tab of the Column Settings pane you can

7 | Evaluating Results256 | NGC Chromatography Systems and ChromLab SoftwareDisplaying Column Performance Results — the Column Performance T

Column Performance AnalysisUser Guide | 257 Width at half height (ml) — displays the width (in ml) of the peak at 50% of the maximum peak height.

2 | The Workspace24 | NGC Chromatography Systems and ChromLab SoftwareThe Method Editor WindowThe Method Editor window enables you to create

7 | Evaluating Results258 | NGC Chromatography Systems and ChromLab Software

User Guide | 2598 Importing and Exporting DataData files can be imported in the Home, Method Editor, or Evaluation window when you choose Import o

8 | Importing and Exporting Data260 | NGC Chromatography Systems and ChromLab SoftwareTo import a method or run 1. In the Home, Method Edi

Importing Unicorn Data FilesUser Guide | 2616. (Optional) Select Open on Import to open in the Evaluation window upon import.Note: This option is

8 | Importing and Exporting Data262 | NGC Chromatography Systems and ChromLab Software3. Click Select Project.4. In the Import Unicorn Dat

Importing BioLogic DuoFlow Data FilesUser Guide | 2639. If it is not already open, select the run to display its chromatogram and trace table in t

8 | Importing and Exporting Data264 | NGC Chromatography Systems and ChromLab Software4. Click Import.ChromLab imports all runs in the dat

Exporting DataUser Guide | 265Exporting Data as an NGC FileYou can export method and run information from the Home, Method Editor, or Evaluation w

8 | Importing and Exporting Data266 | NGC Chromatography Systems and ChromLab SoftwareTo export a method with all of its runs1. With the m

Exporting DataUser Guide | 267To export multiple methods and associated runs 1. In the Home window, select File > Export > Method/Method Run

The Evaluation WindowUser Guide | 2 5The Evaluation WindowThe Evaluation window enables you to view and compare run data, perform peak integration

8 | Importing and Exporting Data268 | NGC Chromatography Systems and ChromLab Software7. Click OK to close the dialog box.To export multip

Exporting DataUser Guide | 269Exporting Run Data as a .csv FileExporting run data to a .csv file enables you to employ various standard file forma

8 | Importing and Exporting Data270 | NGC Chromatography Systems and ChromLab Software2. Clear the checkbox for any run data you do not wa

Exporting DataUser Guide | 271Exporting Diagnostic LogsIn ChromLab, you can export all critical information that Bio-Rad Technical Support require

8 | Importing and Exporting Data272 | NGC Chromatography Systems and ChromLab Software3. (Optional) Clear the Include Runs and Methods in

Exporting DataUser Guide | 273 Do one of the following: Click OK to accept any changes and return to the Export Diagnostic Logs dialog box. Cli

8 | Importing and Exporting Data274 | NGC Chromatography Systems and ChromLab Software

User Guide | 2759 ReportsThe three report formats available in ChromLab™ software make it easy to publish method, run, and analysis data in attrac

9 | Reports276 | NGC Chromatography Systems and ChromLab SoftwareProducing a ReportAfter you generate the data you want to include, you can

Producing a ReportUser Guide | 277Method ReportsNote: The Scout column and Scouting Parameters table appear in the Method report only if the meth

2 | The Workspace26 | NGC Chromatography Systems and ChromLab SoftwareInstrument Control Touch ScreenIn addition to ChromLab software runnin

9 | Reports278 | NGC Chromatography Systems and ChromLab SoftwareAnalysis ReportsNote: You must first perform peak integration analysis in

Saving a ReportUser Guide | 279Saving a ReportTo save a report With the report displayed in the Report Viewer dialog box, click Save and select a

9 | Reports280 | NGC Chromatography Systems and ChromLab Software

User Guide | 281A Database ManagementChromLab™ software stores all data (for example, methods, templates, and runs) in a database using Microsoft

A | Database Management282 | NGC Chromatography Systems and ChromLab SoftwareBack Up the NGC DatabaseYou cannot start the ChromLab Administr

Restore the NGC DatabaseUser Guide | 2833. In the Backup section, click Browse to browse to a location into which to save the NGC backup (.bak) fi

A | Database Management284 | NGC Chromatography Systems and ChromLab Software4. Click Restore.A status bar appears, displaying the restore p

User Guide | 285B Multicolumn PurificationsProtein purification can involve challenging separations of complex mixtures that might not provide suf

B | Multicolumn Purifications286 | NGC Chromatography Systems and ChromLab SoftwareMulticolumn Purification Method TemplatesChromLab™ softwa

Multicolumn Purification Method TemplatesUser Guide | 287Table 7. Multicolumn sequential templates Template Name Template DescriptionAffinity (1

Instrument Control Touch ScreenUser Guide | 2 7Touch Screen Menu CommandsCalibrate — opens the Calibration dialog box, which displays instructions

B | Multicolumn Purifications288 | NGC Chromatography Systems and ChromLab SoftwareAffinity (5 ml) — Step Gradient This method is used to pu

Multicolumn Purification Method TemplatesUser Guide | 289Multicolumn Tandem Purification TemplatesTandem purifications are used when two different

B | Multicolumn Purifications290 | NGC Chromatography Systems and ChromLab Software2-D Affinity (5 ml) > Desalting (50 ml) This method is

Performing Multicolumn Purification ChromatographyUser Guide | 291Performing Multicolumn Purification ChromatographyTo perform multicolumn purific

B | Multicolumn Purifications292 | NGC Chromatography Systems and ChromLab SoftwarePlumbing the NGC Discover Pro System to Use Multicolumn P

Performing Multicolumn Purification ChromatographyUser Guide | 293Multicolumn Tandem 2-D Affinity TemplatesTasks1.Insert a second column switching

B | Multicolumn Purifications294 | NGC Chromatography Systems and ChromLab SoftwarePriming the NGC SystemImportant: Whenever you add or rem

Performing Multicolumn Purification ChromatographyUser Guide | 295Selecting and Saving a TemplateTip: You must save the template as a method bef

B | Multicolumn Purifications296 | NGC Chromatography Systems and ChromLab SoftwareIn the lower-right pane The Overview tab displays a grap

NGC™ Chromatography Systems and ChromLab™ SoftwareUser GuideVersion 3.0

2 | The Workspace28 | NGC Chromatography Systems and ChromLab SoftwareTouch Screen Toolbar CommandsSave — saves in a data file the steps exe

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User Guide | 2 93 System ControlThe ChromLab™ System Control window is the main interface to the NGC™ chromatography system instrument. This inter

3 | System Control30 | NGC Chromatography Systems and ChromLab SoftwareLEGEND1 The menu bar provides quick access to File, View, Man

System Control WindowUser Guide | 3 1File Menu CommandsConnect to System — connects your computer to the NGC system if it is not already connected

3 | System Control32 | NGC Chromatography Systems and ChromLab SoftwareView Menu CommandsShow Chromatogram — displays a chromatogram of the

System Control WindowUser Guide | 3 3Map Fluidic Scheme — opens the Fluidic Scheme Mapping dialog box, which displays the location of two or more

3 | System Control34 | NGC Chromatography Systems and ChromLab SoftwareAutoscale UV Trace — automatically scales the primary y-axis to the t

Chromatogram ViewUser Guide | 3 5Chromatogram ViewA chromatogram shows the real-time acquisition of data during the run as well as its outcome. It

3 | System Control36 | NGC Chromatography Systems and ChromLab SoftwareYou can expand the chromatogram or the fluidic scheme by clicking the

Chromatogram ViewUser Guide | 3 7Table 1. Trace Definitions Trace Module ExplanationUV  1–4 Single-Wavelength UV and Multi-Wavelength UV/Vis Dete

Bio-Rad Technical Support DepartmentThe Bio-Rad Technical Support department in the U.S. is open Monday through Friday, 5:00 AM to 5:00 PM, Pacific Ti

3 | System Control38 | NGC Chromatography Systems and ChromLab SoftwareChanging Chromatogram View SettingsYou can change trace display attri

Chromatogram ViewUser Guide | 3 9When a manual or method run is saved, the new settings are saved with it. When you open the run in the Evaluation

3 | System Control40 | NGC Chromatography Systems and ChromLab SoftwareChanging Trace ColorTo change trace color1. Expand the legend and cli

Chromatogram ViewUser Guide | 4 1Changing the X-Axis Units and ScaleFor runs performed from saved methods, the x-axis unit can be changed to Time

3 | System Control42 | NGC Chromatography Systems and ChromLab SoftwareWhen a single UV absorbance trace is monitored with a single-wave UV

Chromatogram ViewUser Guide | 4 3To add annotations1. Click Annotate on the toolbar and drag the icon onto the chromatogram. The green annotation

3 | System Control44 | NGC Chromatography Systems and ChromLab SoftwareShowing and Hiding Events, Fractions, Errors, and AnnotationsYou can

Chromatogram ViewUser Guide | 4 5When you pause the pointer on a trace, a tooltip displays the real-time reading at the current location in the ch

3 | System Control46 | NGC Chromatography Systems and ChromLab SoftwareFluidic Scheme PaneThe Fluidic Scheme pane graphically displays each

Fluidic Scheme PaneUser Guide | 4 7When pumps are not running, the predicted flow path appears in a lighter color corresponding to the pump flow p

User Guide | iiiTable of ContentsChapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Main NGC Fe

3 | System Control48 | NGC Chromatography Systems and ChromLab SoftwareThe fluidic scheme displays the number of a valve’s active port in br

Fluidic Scheme PaneUser Guide | 4 9ModulesFluidic scheme modules are more fully described in Chapter 2, Components, of the Instrument Guide. For e

3 | System Control50 | NGC Chromatography Systems and ChromLab SoftwareSample PumpThis module dialog box controls the sample pump and displa

Fluidic Scheme PaneUser Guide | 5 1Fluidic Scheme ConfigurationsThe fluidic scheme must be customized to match your instrument hardware setup. In

3 | System Control52 | NGC Chromatography Systems and ChromLab SoftwareSystem pump and inlet BSelect from up to eight different buffers for

Fluidic Scheme PaneUser Guide | 5 3Sample inject valve with sample pumpAutomatically load a specific predetermined sample volume onto a sample loo

3 | System Control54 | NGC Chromatography Systems and ChromLab SoftwareColumns Configuration ExplanationColumn Self-explanatory when column

Fluidic Scheme PaneUser Guide | 5 5Single-wavelength UV with conductivity and pH probeFor selection of a single-wavelength UV detector with conduc

3 | System Control56 | NGC Chromatography Systems and ChromLab SoftwareFraction Collection Configuration ExplanationBioFrac™ fraction collec

Fluidic Scheme PaneUser Guide | 5 7Working with Fluidic SchemesFive sections of the fluidic scheme can be configured based on the available module

iv | NGC Chromatography Systems and ChromLab SoftwareTable of ContentsChromatogram View . . . . . . . . . . . . . . . . . . . . . . . . . . . .

3 | System Control58 | NGC Chromatography Systems and ChromLab Software2. Click an entry in the Fluidic Scheme Selector pane.The right pane

Fluidic Scheme PaneUser Guide | 5 93. Click Edit under the group of modules to change.A spin box displays a number of modules of the type you sele

3 | System Control60 | NGC Chromatography Systems and ChromLab SoftwareFluidic Scheme MappingThe NGC instrument supports up to four inlet va

Fluidic Scheme MappingUser Guide | 6 1 The color and labeling scheme for the Fluidic Scheme Mapping dialog box is as follows:Color Label valveBlue

3 | System Control62 | NGC Chromatography Systems and ChromLab SoftwareTo unmap valves1. Do one of the following: In the System Control win

Fluidic Scheme MappingUser Guide | 6 3To map valves1. In the Fluidic Scheme Mapping dialog box, select a valve to map in either the upper or lower

3 | System Control64 | NGC Chromatography Systems and ChromLab SoftwareCalibrationsThe NGC instrument arrives factory calibrated. For the mo

CalibrationsUser Guide | 6 5Calibrating Flow Rate of PumpsIf the buffer flow seems to be incorrect, you might want to recalibrate the pumps.Import

3 | System Control66 | NGC Chromatography Systems and ChromLab SoftwareCalibrating Pressure SettingsWARNING! You must remove the tubing fr

Verifying Plumbing with the Point-to-Plumb FeatureUser Guide | 6 7Verifying Plumbing with the Point-to-Plumb FeatureThe NGC chromatography system

User Guide | vTable of ContentsDetector Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

3 | System Control68 | NGC Chromatography Systems and ChromLab Softwarepath between them is selected. If the fluidic path includes an NGC mo

System SettingsUser Guide | 6 9System SettingsThe System Settings dialog box enables you to customize your system. From this dialog box you can S

3 | System Control70 | NGC Chromatography Systems and ChromLab SoftwareWhen the delay volume is set, the fraction collector or outlet valves

System SettingsUser Guide | 7 13. Select Synchronize with detector.4. Specify the inner diameter of the tubing in use. Tubing Color Inner Diameter

3 | System Control72 | NGC Chromatography Systems and ChromLab Software7. If your system includes both the BioFrac fraction collector and an

System SettingsUser Guide | 7 3When Control Flow to avoid overpressure is selected, the system lowers the flow rate to avoid exceeding the maximum

3 | System Control74 | NGC Chromatography Systems and ChromLab Software4. (Optional) Specify a minimum flow rate for the system pump and for

System SettingsUser Guide | 7 5To revert trace display settings to their factory defaults1. Select File > System Settings to open the System Se

3 | System Control76 | NGC Chromatography Systems and ChromLab SoftwareDevice Input TabYou can import analog (voltage) signal from up to two

System SettingsUser Guide | 7 74. Choose the Device Input tab.5. By default, the fields in this dialog box are disabled. Select the Enabled checkb

vi | NGC Chromatography Systems and ChromLab SoftwareTable of ContentsMethod Settings Parameters. . . . . . . . . . . . . . . . . . . . . . . .

3 | System Control78 | NGC Chromatography Systems and ChromLab Software6. (Optional) Select the Enabled checkbox for SIM 2 and repeat steps

System SettingsUser Guide | 7 95. By default, the fields in this dialog box are disabled. Select the Enabled checkbox for Signal Output A and prov

3 | System Control80 | NGC Chromatography Systems and ChromLab SoftwareAir Sensors TabThe system supports up to two air sensor modules and u

System SettingsUser Guide | 8 13. To activate one or more air sensors, select its checkbox and from the dropdown list choose either Sample (to det

3 | System Control82 | NGC Chromatography Systems and ChromLab SoftwareEmail Notifications TabYou can connect ChromLab to your outgoing emai

System SettingsUser Guide | 8 33. Select Enable Email Notification.Note: If you have not set up a valid SMTP connection for ChromLab, an error me

3 | System Control84 | NGC Chromatography Systems and ChromLab SoftwareAll added users receive an email from DoNotReply_NGC notifying them t

System SettingsUser Guide | 8 5System Name TabYou can specify a unique name for the NGC instrument. ChromLab uses this name to identify the system

3 | System Control86 | NGC Chromatography Systems and ChromLab SoftwareSystem InformationThe System Information dialog box enables you to vi

System InformationUser Guide | 8 7Detector TabThis tab displays details about UV or UV/Vis lamp usage, from which you can determine whether the la

User Guide | viiTable of ContentsCollecting Fractions in Run Queues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174Collecting

3 | System Control88 | NGC Chromatography Systems and ChromLab SoftwareTo reset the lamp hours On the Detector tab, click Reset Lamp Time.N

PreferencesUser Guide | 8 9PreferencesThe Preference dialog box enables you to select pressure units for all system and software pressure values.

3 | System Control90 | NGC Chromatography Systems and ChromLab SoftwareTo connect ChromLab to an email server 1. In ChromLab select File >

PreferencesUser Guide | 9 1 Authentication — by default, Authentication is disabled. If your site requires account authentication, select this ch

3 | System Control92 | NGC Chromatography Systems and ChromLab Software

User Guide | 9 34 Performing a Manual RunThe NGC™ chromatography system instrument operates in two modes: manual mode and automated, programmed me

4 | Performing a Manual Run94 | NGC Chromatography Systems and ChromLab SoftwarePreparing the SystemSystem preparation consists of priming

Preparing the SystemUser Guide | 9 55. Prime the system. (See Priming and Purging the Systems in Chapter 3 of the NGC Chromatography Systems and C

4 | Performing a Manual Run96 | NGC Chromatography Systems and ChromLab Software4. In the System Pump dialog box:a. Set Flow Rate to 10 ml

Preparing the SystemUser Guide | 9 7To purge the buffer blending valve1. In the fluidic scheme, double-click the System Pump module to open its di