Bio-Rad Human MMP and TIMP Assays - Manual (Page 21)

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Use calibrated pipets and pipet carefully, avoiding bubbles
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Pay close attention to vortexing, shaking, and incubation instructions.
Deviation from the protocol may result in low assay signal and
assay variability
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Assay incubations are carried out on a shaker at 850 ± 50 rpm at room
temperature (RT). Cover the plate with sealing tape and protect from
light with aluminum foil
Table 8. Summary of wash options and protocols. After each assay step, select the
appropriate Bio-Plex Pro ™ wash station program or perform the appropriate manual wash
step as summarized below.
Add Coupled Beads, Samples, Standards, Blank,
and Controls
1. Cover unused wells of the assay plate with sealing tape.
2. Vortex the diluted (1x) beads for 30 sec at medium speed. Pour into
a reagent reservoir and transfer 50 µl to each well of the assay plate.
Tip: A multichannel pipet is highly recommended for ease of use
and efficiency.
3. Wash the plate two times with 100 µl Bio-Plex ® wash buffer per
well, using the wash method of choice.
4. Vortex the diluted samples, standards, blank, and controls at medium
speed for 5 sec. Transfer 50 µl of each to the appropriate well of the
assay plate, changing the pipet tip after every volume transfer.
5. Cover plate with a new sheet of sealing tape and protect from light
with aluminum foil. Incubate on shaker at 850 ± 50 rpm for 1 hr at RT.
Note: Be consistent with this incubation time and shaker setting for
optimal assay performance and reproducibility.
Handheld Magnet or
Bio-Plex Pro Wash Station Vacuum Manifold
Assay Step Magnetic Program Manual Wash Steps
Add beads to plate MAG x2 2 x 100 μl
Sample incubation
Detection Ab incubation
MAG x3 3 x 100 μl
SA-PE incubation