Bio-Rad Bio-Plex Pro™ Human Th17 Cytokine Assays - Manual (Page 33)

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Possible Causes
Low Bead Count
Miscalculation of bead dilution
Beads clumped in multiplex
bead stock tube
Vacuum on for too long when
aspirating buffer from wells
Assay plate not shaken enough
during incubation steps and prior
to reading
Reader is clogged
Incorrect needle height of the
reader
Low Signal or Poor Sensitivity
Standards reconstituted incorrectly
Detection antibody or
streptavidin-PE diluted incorrectly
Possible Solutions
Check your calculations and be
careful to add the correct volumes
Vortex for 30 sec at medium speed
before aliquoting beads.
Do not apply vacuum to the filter
plate for longer than 10 sec after the
buffer is completely drained from
each well.
Shake the plate at 850 ±50 rpm
during incubation steps and for
30 sec immediately before reading
the plate.
Refer to the troubleshooting guide
in the Bio-Plex system hardware
instruction manual (bulletin
#10005042).
Adjust the needle height to coincide
with the plate type provided in the kit.
Follow the cytokine standard
instructions carefully.
Check your calculations and be
careful to add the correct volumes.