Bio-Rad Bio-Plex Pro Human Angiogenesis Reagent and Diluent Kits - Manual (Page 29)

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Possible Causes Possible Solutions
High Background Signal
Incorrect buffer was used
(for example, assay buffer
used to dilute standards) Use sample matrix or serum
standard diluent to dilute
the standards.
Spiked “0 pg/ml” wells by mistake Be careful when spiking standards.
Do not add any antigens in the
0 pg/ml (blank) point.
Streptavidin-PE incubated
too long Follow the procedure incubation
time.
Poor Recovery
Expired Bio-Plex reagents were
used Check that reagents have not
expired. Use new or unexpired
components.
Incorrect amounts of components
were added Check your calculations and be
careful to add the correct volumes.
Microplate shaker set to an
incorrect speed 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.
Pipetting technique Pipet carefully when adding
standards, samples, detection
antibodies, and streptavidin-PE,
especially when using a
multichannel pipet. Use a calibrated
pipet. Change pipet tip after every
volume transfer.