Bio-Rad Bio-Plex Pro™ Human Chemokine Assays - Manual (Page 19)

3. Vortex the 20x stock of 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 requires 2.5 μl of the 20x 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 20x 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 20x stock (includes 20% excess volume)
Table 6. Premixed panel or one singleplex assay.
Table 7. Mixing singleplex assays.
17
# of Wells 20x Beads, µl Assay Buffer, µl Total Volume, µl
96 288 5,472 5,760
48 144 2,736 2,880
20x Beads, µl 20x Beads, µl
# of Wells Singleplex #1 Singleplex #2 Assay Buffer, µl Total Volume, µl
96 288 288 5,184 5,760
48 144 144 2,592 2,880