Bio-Rad Nonmagnetic Beads and Related Reagents - Manual (Page 3)

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Section 1
Introduction
The Bio-Plex Pro Magnetic COOH Beads and Bio-Plex COOH Beads are
suited for coupling to proteins or nucleic acids. Bio-Plex Pro Magnetic Beads
are identical to MagPlex
® Microspheres, and Bio-Plex COOH Beads are
identical to Microplex Microspheres, both developed by Luminex corporation.
This instruction manual describes the coupling procedures.
The Bio-Plex Amine Coupling Kit (catalog #171-406001) contains all of the
buffers necessary to covalently couple 6–150 kD proteins to 6.5 µm Bio-Plex
Pro Magnetic COOH beads (magnetic, carboxylated) or 5.5 µm Bio-Plex
COOH beads (nonmagnetic, carboxylated) in under three hours. Coupling is
achieved via carbodiimide reactions involving the primary amino groups on the
protein and the carboxyl functional groups on the bead surface. The covalent
attachment is permanent, leaving no unbound protein after cleanup, even after
months of storage. The contents of the kit are sufficient for 30 coupling
reactions. Each coupling reaction requires 1.25 x 10
6 carboxylated beads (1x
scale). For larger scale coupling reactions, the volume of the buffers used in
each step can be proportionally increased to a maximum 10x scale. The
protein-coupled beads can then be used in multiplex protein-protein binding
studies or in the development of multiplex assays that can be analyzed with the
Bio-Plex suspension array system. The bead yield per coupling reaction is
approximately 80%, enough protein-coupled beads for four 96-well microtiter
plates using 2,500 beads per well.
For nucleic acid oligonucleotide coupling, refer to this manual for ordering
information and buffer recipes. The amine coupling kit contains buffers used for
coupling proteins only.
The coupling accessories for multiplex assay development complement the
Bio-Plex assay products and associated kits. For a current listing of Bio-Plex
products,visit us on the Web at www.bio-rad.com/bio-plex.