Bio-Rad Human MMP and TIMP Assays - Manual (Page 20)

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3. Vortex the 10x 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 5 μl of the 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 10x 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 6. Premixed panel or one singleplex assay.
Table 7. Mixing singleplex assays.
8. Run Assay
Considerations
n
Bring all assay components and samples to room temperature
before use
# of Wells 10x Beads, µl Assay Buffer, µl Total Volume, µl
96 575 5,175 5,750
48 288 2,587 2,875
Singleplex #1 Singleplex #2
# of Wells 10x Beads, µl 10x Beads, µl Assay Buffer, µl Total Volume, µl
96 575 575 4,600 5,750
48 288 288 2,300 2,876