Bio-Rad Nonmagnetic Beads and Related Reagents - Manual (Page 33)

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Section 12
Troubleshooting Guide
This troubleshooting guide addresses problems that may be encountered
during a protein coupling reaction. If you experience any of the problems listed
below, review the possible causes and solutions provided. Contact Luminex
Tech Support for technical issues related to coupling of oligonucleotides
(source: http://www.luminexcorp.com/support/tech_tips.html).
Possible Causes Possible Solutions
Low MFI signal in validation procedure
EDAC and S-NHS may have
expired
Purified protein was not used
immediately during the conjugation
procedure or was not kept on ice
S-NHS was not added to the
beads immediately after adding
EDAC
Wrong antibody was used in the
validation procedure
Use fresh EDAC and S-NHS for
each conjugation
Keep the protein on ice during the
conjugation procedure
Add S-NHS to the beads
immediately after adding EDAC
Ensure that the correct species-
specific antibody is being used for
validation (if an antibody is coupled)