Bio-Rad Bio-Plex® Assay Builder - Manual (Page 14)

Section 9
Safety Considerations
Eye protection and gloves are recommended while using this product.
Consult the MSDS for additional information.
Human Source Material. Treat As Potentially Infectious.
The lysates provided with Bio-Plex phosphoprotein and total target assay
contain components of human origin. The components are known to
contain an agent that requires handling at Biosafety Level 2 containment
[US Government Publication: Biosafety in Microbiological and Biomedical
Laboratories (CDC, 1999)]. These agents have been associated with
human disease. These components have not been screened for hepatitis
B, human immunodeficiency viruses, or other adventitious agents.
Handle Bio-Plex phosphoprotein positive and negative controls as
potentially biohazardous material under at least Biosafety Level 2
containment.
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Contamination with wash buffer
during wash steps
Microplate shaker set to an
incorrect speed
Cellular debris not cleared from
lysate
Bead resuspension buffer not used
after streptavidin-PE incubation
High Background Signal
(From Both Lysates Provided with
the Assays and Experiment Lysates)
Vacuum pressure too low,
resulting in residue in wells
Wash steps performed incorrectly
Streptavidin-PE incubation
step too long
During the wash steps, do not splash
wash buffer from one well to another.
Filter the wells completely so that no
residual volume remains. Also, be sure
that the microplate shaker setting is not
too high. Reduce the microplate shaker
speed to minimize splashing
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
Be sure to remove cellular debris by
centrifugation as directed. Avoid
disturbing the pellet while collecting the
supernatant
Wash 3 times with bead resuspension
buffer after streptavidin-PE incubation
as described in the assay instructions
Use recommended filter plate vacuum
apparatus with proper vacuum pressure
setting
Perform washes as described in the
assay instructions
Check suggested incubation times for
appropriate steps of the assay. Follow
the suggested time for incubation
Possible Causes Possible Solutions