Bio-Rad Bio-Plex Pro™ Human Isotyping Assays - Manual (Page 26)

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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.
b. For the vial of quality controls supplied, format the appropriate
wells as controls and enter descriptions, but leave the
concentrations blank. Alternatively, the quality 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 and enter sample information and the
appropriate dilution factor.
7. Click Run Protocol and confirm that the 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 and 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.
c. 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.