Bio-Rad Bio-Plex® Precision Pro™ Human Cytokine Assays User Manual
Page 14
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Assay Procedure
Bring all buffers to room temperature. Avoid bubbles when pipetting.
Assay Key – The following terms are repeated throughout the assay
procedure. Refer to these detailed instructions when wash, incubate, and
vacuum-filter are shown in bold.
1. Equilibrate the diluted standards, samples, and controls at room
temperature for 20 min prior to use.
2. Prewet and block the desired number of wells in a 96-well filter plate
with 100 µl of assay buffer and
vacuum-filter. If fewer than 96 wells
are required, mark the plate to identify the unused wells for later use
and cover the unused wells with sealing tape.
3. Vortex the coupled magnetic beads (1x) for 15–20 sec at medium
speed. Add 50 µl to each well and immediately
vacuum-filter.
4.
Wash twice.
5. Gently vortex the diluted standards, controls, and samples for 1–3
sec. Add 50 µl of standard, control, or sample to each well, changing
the pipet tip after every volume transfer.
Incubate for 1 hr.
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Gently cover the filter plate with a new sheet of sealing tape. Place
the filter plate on a microplate shaker and then cover with aluminum
foil. Shake the filter plate at room temperature at 1,100 rpm for
30 sec, then at 300 rpm for the specified incubation time.