Bio-Rad Bio-Plex Pro™ Human Cancer Biomarker Assays - Manual (Page 27)

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5. Enter Controls Info into Bio-Plex Manager.
a. For user-specified controls, select an analyte from the dropdown
menu, then enter a description and concentration. Repeat for
each additional analyte in the assay.
b. For the quality controls supplied in premixed cancer
biomarker kits, format the appropriate wells as controls, enter
descriptions, but leave the concentrations blank. Alternatively,
the quality controls can be formatted as samples with clear
descriptions such as “quality control high” and “quality control
low.” In any case, the expected control ranges provided on the
product data sheet are not entered into Bio-Plex Manager
software version 6.1 and earlier.
6. Enter Sample Info — enter sample information and the appropriate
dilution
factor.
7. Run Protocol — confirm that the assay settings are correct.
a. Human cancer biomarker assays are run at the default setting in
Bio-Plex Manager: low RP1 target (low PMT).
b. Confirm that data acquisition is set to 50 beads per region.
In Advanced Settings, confirm that the bead map is set to
100 region, the sample size is set to 50 µl, and the doublet
discriminator (DD) gates are set to 5,000 (Low) and 25,000 (High).
In Bio-Plex Manager software versions 4.0, 4.1, 4.1.1, and 5.0,
check Override Gates and set the DD gate values as indicated.
Select Start, name and save the .rbx file, and begin data
acquisition. The Run Protocol pop-up screen will appear.
Click Eject/Retract to eject the plate carrier.