Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 22)

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3. Vortex the stock coupled beads at medium speed for 30 sec.
Carefully open the cap and pipet any liquid trapped in the cap back
into the tube. This is important to ensure maximum bead recovery.
Do not centrifuge the vial; doing so will cause the beads to pellet.
4. Dilute coupled beads to 1x by pipetting the required volume into the 15 ml
tube. Vortex.
Each well of the assay plate requires either 2.5 µl (20x stock) or 5.0 µl (10x
stock) adjusted to a final volume of 50 µl in assay buffer.
5. Protect the beads from light with aluminum foil. Equilibrate to room
temperature prior to use.
Note: To minimize volume loss, use a 200–300 μl capacity pipet
to remove beads from the stock tube. If necessary, perform the
volume transfer in two steps. Do not use a 1,000 μl capacity pipet
and/or wide bore pipet tip.
Preparing 1x coupled beads from 10x stock (includes 20% excess volume)
Table 10. Premixed panel or one singleplex assay.
Table 11. Mixing two singleplex assays or a premixed panel + singleplex assay.
# of Wells 10x Beads, µl Assay Buffer, µl Total Volume, µl
96 575 5,175 5,750
48 288 2,587 2,875
10x Beads, µl 10x Beads, µl
# of Wells Singleplex #1 Singleplex #2 Assay Buffer, µl Total Volume, µl
96 575 575 4,600 5,750
48 288 288 2,300 2,876