Dip and Readâ¢ Residual Protein A Detection Kit - ForteBio

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Dip and Read Residual Protein A Detection Kitb Under Quantitation Sensors click Add…cEnter Residual Protein A as the new biosensor name andclick OK.d Click Close. This sensor type is now available to all users ofthe instrument.Standard Assay Protocol1 At least 40 minutes before running the assay, turn on theOctet instrument and associated computer. It is critical thatthe lamp warms up for at least 40 minutes prior to the firstmeasurement.2 Start the Data Acquisition software (version 6.1 or later).3 From the Experiment Wizard (ctrl+N) choose the AdvancedQuantitation option.4 In tab 1:aClick the Modify button and choose the Residual Protein ADetection method from the list on the left. The settings inthis list are default settings. To modify the settings use theup or down arrows.b Select the Sample Plate radio button and right-click on theSample plate map to open the contextual menu to definethe location of the samples, controls, references, and calibratorsin the sample plate (i.e., the microplate used duringthe offline biosensor incubation step).cEnter Sample IDs, concentrations and dilution factors asdesired in the Sample Plate table.d Select the Reagent Plate radio button and right-click onthe Reagent plate map to open the contextual menu todefine the location of the buffer (Sample Diluent+PEG)and the detection reagent (diluted Detection Reagent).5 In tab 2:aDefine the locations of the biosensors. Default locationsare shown automatically. Locations can be changed byhighlighting the biosensors and using the Remove, Fill orFill Plate buttons.7 Start the assay by clicking the Go button. The default settingswill ensure the data, method and runtime charts are savedautomatically.D: Data Analysis and Report Generation1 Start the Data Analysis software (version 6.1 or later).2 In tab 1:aLoad data by locating the data folder in the lower explorertree, right click on it, and choose Load Folder.b Select data to analyze by clicking on the data folder in theQuantitation section of the Loaded Data tree. Multiple datasets can be selected for analysis. All selected data sets willbe analyzed as a group.cOnce the data is loaded, sample ID, group, concentrationand dilution factors can be modified in the table ifneeded.d If needed, use the “Reference Subtraction” option to subtractreference well values from the remaining sample wells.3 In tab 2:aSelect the desired standard curve.b Select R equilibrium as the binding rate equation. Thisequation will fit the binding curve generated during theexperiment and calculate a response at equilibrium as theoutput signal.cClick Calculate Binding Rate. Results will be displayed automaticallyin the table.4 Click the Save Report button or select File > Save Report togenerate a Microsoft® Excel formatted report.E: Representative DataFigure 1 shows representative raw data and standard curve generatedusing the Dip and Read Residual Protein A Detection Kit.b Choose Residual Protein A from the Sensor Type dropdownmenu.cIf desired, add biosensor information in the table on theright.6 in tab 3:aEnter the location and folder name into which the datashould be saved.b Enter a delay if needed to complete the 5-minute minimumsample equilibration time.4
Technical Note 18Binding, nm2.62.42.22.01.81.61.41.21.00.80.60.40.20.00 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300Time (sec)Recombinant Protein ACalibrator Concentration25 ng/mL10 ng/mL5 ng/mL2.5 ng/mL1.0 ng/mL0.5 ng/mL0.25 ng/mL0.1 ng/mL0 ng/mL3.02.82.62.42.22.0Binding, nm1.81.61.41.21.00.80.60.40.20.00 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25Concentration (ng/mL)Figure 1: Typical binding data and resulting standard curve from Recombinant Protein A calibrator levels at 25, 10, 5, 2.5, 1, 0.5, 0.25, and 0.1 ng/mL.5
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