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

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Possible Causes
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
Poor Recovery
Expired Bio-Plex reagents
were used
Incorrect amounts of components
were added
Microplate shaker set to an
incorrect speed
High end saturation of the
standard curve
Quality controls do not fall within
expected ranges.
Possible Solutions
Use isotyping diluent to dilute
standards.
Do not add any antigens to the
blank wells.
Follow the procedure incubation
time precisely.
Check that reagents have not expired.
Use new or nonexpired components.
Check your calculations and be
careful to add the correct volumes.
Check the microplate shaker speed
and use the recommended setting.
Setting the speed too high may
cause splashing and contamination.
Use the recommended plate shaker.
Make sure that correct shaker speed
and incubation times are used.
Remove S1 for data analysis if needed.
Make sure that the vial of controls
is reconstituted at the same time as
standards and in the same diluent.
Incubate for precisely 30 min.