Bio-Rad Bio-Plex Pro™ Human Chemokine Assays - Manual (Page 34)

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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 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. Setting the speed too low may
cause low assay signal and false plateau or
high end saturation of standard curves.
Make sure that correct shaker speed and
incubation times are used. Remove S1 for
data analysis if needed.
Make sure that the controls are
reconstituted at the same time as standards
and in the same diuent (standard diluent
HB). Incubate for precisely 30 min.