Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 18)

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6. Run the Assay
Considerations
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Bring all assay components and samples to room temperature before use
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Use calibrated pipets and pipet carefully, avoiding bubbles. Use new
pipet tips for every volume transfer
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Pay close attention to vortexing, shaking, and incubation instructions.
Deviation from protocol may result in low assay signal and assay variability
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Assay incubations are carried out in the dark on a shaker at 850 ± 50 rpm.
Cover the plate with a plate seal and protect from light with aluminum foil
Table 6. 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.
Considerations When Using a Vacuum Manifold
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After each incubation, place the filter plate on a calibrated vacuum
apparatus and remove the liquid by vacuum filtration
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To wash, add 100 μl wash buffer to each well and remove the liquid as
before. Ensure that all wells are exposed to the vacuum
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Thoroughly blot the bottom of the filter plate with a clean paper towel
between each vacuum step to prevent cross contamination
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Place the assay plate on the plastic plate holder/tray as needed
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Before each incubation, gently cover the plate with a new plate seal.
Avoid pressing down on the wells to prevent leaking from the bottom
Bio-Plex Pro or Bio-Plex Pro II Handheld Magnet or
Pro II Wash Station Wash Station Vacuum Manifold
Assay Step Magnetic Program Vacuum Program Manual Wash Steps
Sample incubation
MAG x3 VAC x3 3 x 100 μl
SA-PE incubation