Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 30)

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Possible Causes
Low Bead Count
Vacuum on for too long when
aspirating buffer from wells
Reader is clogged
Low Signal or Poor Sensitivity
Standards reconstituted
incorrectly
Detection antibody or
streptavidin-PE prepared incorrectly
High Background Signal
Incorrect buffer was used
(for example, assay buffer used
to dilute standards)
Accidentally spiked blank wells
Detection antibodies or
streptavidin-PE incubated too long
Possible Solutions
Do not apply vacuum to the filter
plate for longer than 10 sec after the
buffer is completely drained from
each well.
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.
Use standard diluent to dilute
standards.
Do not add any antigens to the
blank wells.
Follow the procedure incubation
time precisely.