Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 40)

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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
Possible Solutions
Use standard diluent or diluent
similar to final sample matrix 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.