Bio-Rad Bio-Plex Pro Human Angiogenesis Reagent and Diluent Kits - Manual (Page 28)

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Possible Causes Possible Solutions
Low Bead Count
Miscalculation of bead dilution Check your calculations and be
careful to add the correct volumes.
Beads clumped in multiplex
bead stock tube Vortex for 15–20 sec at medium
speed before aliquoting beads.
Vacuum on for too long when
aspirating buffer from wells Do not apply vacuum to the filter
plate for longer than 10 sec after the
buffer is completely drained from
each well.
Did not shake filter plate enough
before incubation steps and prior
to reading Shake the filter plate at 1,100 rpm
for 30 sec before incubation steps
and immediately before reading
the plate.
Reader is clogged Refer to the troubleshooting guide
in the Bio-Plex hardware
instruction manual.
Low Signal or Poor Sensitivity
Standards reconstituted incorrectly Follow the standard preparation
instructions carefully (section 7).
Detection antibody or
streptavidin-PE diluted incorrectly Check your calculations and be
careful to add the correct volumes.