Bio-Rad Bio-Plex Pro™ Magnetic Cell Signaling Assays - Manual (Page 17)

15
Setting up a Vacuum Manifold
Calibrate the vacuum manifold by placing a standard 96-well flat bottom
plate on the unit and adjusting the pressure to between –1 and –3" Hg. In
general, 200 μl liquid should take 5–6 sec to clear the well. For detailed
instructions, refer to bulletin 10005042.
Using a Vacuum Manifold
n
After each incubation, place the filter plate on a calibrated vacuum
apparatus and remove the liquid by vacuum filtration
n
To wash, add 200 μl wash buffer to each well and remove the liquid as
before. Ensure that all wells are exposed to the vacuum
n
Thoroughly blot the bottom of the filter plate with a clean paper towel
between each vacuum step to prevent cross contamination
n
Place the assay plate on the plastic plate holder/tray as needed
n
Before each incubation, gently cover the plate with a new sheet of
sealing tape. Avoid pressing down over the wells to prevent leaking
from the bottom
Bio-Plex Pro Wash Station/Handheld Magnet/Vacuum Manifold*
Wash Program Settings and Manual Wash Steps
Assay Step # of Washes Volume, µl Magnetic Soak, min
1. Add beads to plate 2 200 1
2. Sample incubation 3 200 1
3. Detection Ab incubation 3 200 1
4. SA-PE incubation 3 200 1
* No magnetic soak for vacuum filtration.
Table 5. Summary of wash steps and settings. After each assay incubation step, perform the
appropriate wash step as shown below.