Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 23)

21
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.
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.
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.