Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 34)

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7. Click Run Protocol and confirm that the assay settings are correct.
a. Refer to Table 24 for the recommended RP1 (PMT) setting.
Protocols using alternative PMT settings should be validated
by the end user, for example when mixing diabetes assays with
cytokine assays.
b. Confirm 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 DD gates are
set to 5,000 (Low) and 25,000 (High). In Bio-Plex Manager
software versions 4.0, 4.1, and 4.1.1, 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.
2. Click Run Protocol — on the pop-up screen, select Load Plate and
click OK to start acquiring data.
3. Use the Wash Between Plates command after every plate run
to reduce the possibility of clogging the instrument.
4. If acquiring data from more than one plate, empty the waste bottle
and refill the sheath bottle after each plate (if HTF are not present).
Select Wash Between Plates and follow the instructions. Then
repeat the Prepare Protocol and Acquire Data instructions.
5. When data acquisition is complete, select Shut Down and
follow the instructions.