Bio-Rad Bio-Plex Pro™ RBM Canine Kidney Toxicity Assays - Manual (Page 21)

c. Optional: enter the lot number of the vial of standards into the
Standard Lot box and click Save.
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 and enter sample information and the
appropriate dilution factor.
7. Click Run Protocol and confirm that the settings follow Table 9.
Table 9. 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.
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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.