Each assay tube is acquired on the flow cytometer using a CBA acquisition template. Acquisition template may be downloaded via the Internet from www. bdbiosciences.com/pharmingen/CBA//Dual-Laser.pdf. This template assures that each collected sample file contains approx 300 events for each capture bead species. To facilitate analysis of data files using the BD CBA software a numeric suffix is added to each file that corresponds to the assay tube number (i.e., tube no. 1 containing 0 pg/mL could be saved as RV032595.001). The acquired FACS files are saved and then analyzed using the BD CBA software. The outputs of the analysis are the calibration curves for the six analytes (Fig. 2) and tabulated concentrations of the six cytokines for each sample.
3.2. Soluble Human Protein Flex Set Bead Assay
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