Luminex xMAP Antibody Coupling Kit - Manual (Page 15)

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16. Wash the microspheres.
A. Place the reaction tube with microspheres into the magnetic separator for 1-2
minutes.
B. With the tube still positioned in the magnetic separator, remove the supernatant
with a transfer pipette.
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
disperse the microspheres.
17. Repeat Step 16 twice, for a total of three washes.
18. Place the reaction tube with microspheres into the magnetic separator for 1-2
minutes and, with the tube still positioned in the magnetic separator, remove the
supernatant with a transfer pipette.
19. Calculate volume of antibody to be used in the reaction.
• Typically, a range of 2-5 μg of antibody per 1x10
6
microspheres is a good
starting point, if performing the coupling reaction for the first time.
Example Calculation:
If coupling 5 million microspheres, with an antibody at a concentration of 5 mg/mL,
and using the suggested starting point of 5 μg of antibody per 1x10
6
micro-
spheres: