Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 25)

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5. Click Enter Controls Info.
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 kit controls supplied, format the appropriate wells as
controls and enter descriptions, but leave the concentrations blank.
Alternatively, the controls can be formatted as samples with clear
descriptions such as “quality control.” 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. Click Enter Sample Info — enter sample information and the
appropriate dilution factor.
7. Click Run Protocol — confirm that the assay settings follow Table 11.
Table 11. Read the plate using the appropriate instrument settings.
a. Confirm that data acquisition is set to 50 beads per region.
b. In Bio-Plex Manager software prior to 6.1, go to Advanced Settings,
confirm that the bead map is set to 100 region, the sample size is set
to 50 µl, and the 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.
Acquire Data
1. Shake the assay plate at 850 ± 50 rpm for 30 sec, and visually inspect
the plate to ensure that the assay wells are filled with buffer. Slowly
remove the sealing tape and any plate cover before placing
the plate on the plate carrier.
Instrument RP1 (PMT) DD Gates Bead Events
Bio-Plex 100, 200* Low 5,000 (low), 25,000 (high) 50
Bio-Plex 3D* Standard Select MagPlex beads 50
Bio-Plex ® MAGPIX ™ * N/A, use default instrument settings
* A similar Luminex-based system may be used.