Luminex xMAP Antibody Coupling Kit - Manual (Page 13)

Page 9 of 20
4. Dispense the desired amount of microspheres from the stock vial into one of the
microcentrifuge tubes (“reaction tube”) provided with the kit.
Example Calculation:
If coupling 5 million microspheres, with microsphere stock concentration of
12.5x10
6
microspheres:
IMPORTANT: This kit is designed for a maximum of 12.5x10
6
microspheres per
reaction tube.
5. Wash the microspheres.
A. Place the reaction tube with microspheres into the magnetic separator for 1-2
minutes.
NOTE: If performing multiple reactions simultaneously, a microcentrifuge may be
used, in place of a magnetic separator, to pellet the microspheres during wash
steps. Beads can be pelleted by microcentrifugation at
≥ 8000 x g for 1-2 minutes.
B. With the tube still positioned in the magnetic separator, remove the supernatant
with transfer pipette.
NOTE: For your convenience, twenty disposable transfer pipettes have been
included for the removal of supernatant during the various wash steps in this pro-
tocol. If using one pipette for the removal of Activation Buffer and one for the
removal of Wash Buffer, the kit should have enough to perform up to ten individual
reactions. (i.e. 2/rxn) Special care should be taken to avoid cross contamination,
when performing multiple reactions simultaneously.
C. Add 500 μL of Activation Buffer into the reaction tube.
D. Vortex the reaction tube for 10 seconds and then sonicate for 10 seconds to dis-
perse the microspheres.
6. Repeat Step 5 once for a total of two washes.