Bio-Rad Bio-Plex Pro™ RBM Canine Kidney Toxicity Assays User Manual
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Possible Causes
High Intra-Assay CV
Improper pipetting technique
Reagents and assay components
not equilibrated to room
temperature prior to pipetting
Contamination with buffer
during wash steps
Slow pipetting of samples and
reagents across the plate
Low Bead Count
Beads clumped in multiplex
bead stock tube
Possible Solutions
Pipet carefully when adding
standards, samples, detection
antibodies, and streptavidin-PE,
especially when using a multichannel
pipet. Use a calibrated pipet. Change
pipet tip after every volume transfer.
All reagents and assay components
should be equilibrated to room
temperature prior to pipetting.
During the wash steps, be careful
not to splash buffer from one well
to another. Be sure that the wells
are filtered completely and that no
residual volume remains. Ensure
that the microplate shaker setting is
not too high. Reduce the microplate
shaker speed to minimize splashing.
Sample pipetting across the entire
plate should take less than 4 min.
Reagent pipetting across the entire
plate should take less than 1 min.
Vortex for 30 sec at medium speed
before aliquoting beads.