Bio-Rad Rat Diabetes Assays - Manual (Page 38)

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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 wash buffer
during wash steps
Slow pipetting of samples and
reagents across the plate
Bio-Plex Wash Station:
insufficient washing due to
clogged pins
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 wash 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.
Clean dispensing pins with the thicker
of the 2 cleaning needles provided
with washer. Perform regular rinses to
minimize salt build up.