Acquire data – Bio-Rad Rat Diabetes Assays User Manual

Page 34

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7. Click Run Protocol and confirm that the assay settings are correct.

a. The Bio-Plex Pro diabetes assays were developed on the high

RP1 (high PMT) setting using the Bio-Plex 200 system. 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.

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