Bio-Rad Bio-Plex Pro™ TGF-β Assays - Manual (Page 35)

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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
Filter plate not shaken enough
before incubation steps and prior
to reading
Reader is clogged
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 filter plate at
850 ± 50 rpm for 30 sec before
incubation steps and immediately
before reading the plate.
Refer to the troubleshooting guide
in the Bio-Plex system hardware
instruction manual (bulletin
#10005042).
Follow the standard preparation
instructions carefully.
Check your calculations and be
careful to add the correct volumes.