Bio-Rad Nonmagnetic Beads and Related Reagents - Manual (Page 16)

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Section 7
Protein Coupling
Bring all the buffers to room temperature prior to use. Protect the beads
from light as much as possible by covering the tubes with aluminum foil
during the procedure.
Remove EDAC and S-NHS from the –20ºC freezer and store them in a
desiccator at room temperature for approximately 1 hr prior to their use.
The bottles of EDAC and S–NHS should be discarded after five uses.
7.1 Sample Protocol Using Bio-Plex Pro Magnetic COOH Beads
Notes:
a. Beads must be completely protected from light throughout this
procedure.
b. The stock bead concentration is 1.25 x 10 7 beads/ml.
c. 1x scale = 1.25 x 10 6 beads.
d. Volume stock beads required (ml) = [(1.25 Ч 10 6 ) Ч (scale)] ч [1.25 Ч
107 bead/ml]
Procedure:
1. Vortex the stock uncoupled beads at speed 7 for 30 sec, then
sonicate for 15 sec.
2. For a 1x scale coupling reaction, transfer 100 µL of monodisperse
COOH beads (1.25 x 10 6 beads) to one of the coupling reaction
tubes provided with the kit.
3. Place the tube into a magnetic separator and allow separation to
occur for 30 to 60 seconds.
4. With the tube still positioned in the magnetic separator, remove the
supernatant. Take care not to disturb the beads.
5. Remove the tube from the magnetic separator and resuspend the
beads in 100 µL bead wash buffer by vortex for approximately 30 seconds.