Run the assay, Run the assay 14, Considerations – Bio-Rad Bio-Plex Pro™ RBM Canine Kidney Toxicity Assays User Manual
Page 16: Considerations when using a vacuum manifold
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14
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
n
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 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
n
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 over 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