Bio-Rad Human MMP and TIMP Assays - Manual (Page 18)

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7. Prepare Coupled Beads
1. Use Table 6 or the Calculation Worksheet on page 34 to calculate the
volume of coupled beads and assay buffer needed.
2. Add the required volume of Bio-Plex ® assay buffer to a 15 ml
polypropylene tube.
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.
Table 6. Preparing 1x coupled beads from 20x stock (includes 20% excess volume).
# of Wells 20x Beads, µl Assay Buffer, µl Total Volume, µl
96 288 5,472 5,760
48 144 2,736 2,880