Luminex xMAP Antibody Coupling Kit - Manual (Page 12)

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Instructions
General Workflow
Coupling Protocol
The following is the detailed step-by-step process for coupling antibodies (or similar
proteins) to carboxylated magnetic microspheres. An abridged quick reference guide
(part number 89-30000-00-361) can be found online at http://www.luminexcorp.com/
downloads.
IMPORTANT: Protect photosensitive microspheres from light whenever possible
throughout this entire procedure.
1. Remove kit and all reagents from the refrigerator and allow them to equilibrate to
room temperature for 20-30 minutes.
2. While the reagents equilibrate to room temperature, calculate and note the required
volumes for Decision Point steps 4, 8, 11, 13, 19, and 20; for each reaction being
performed.
3. Resuspend the stock microspheres.
• If using a 1mL stock microsphere vial, vortex the stock microsphere vial for 10
seconds and then sonicate for 10 seconds to disperse the microspheres.
Alternatively, the microsphere vials can be rotated on a rotator for 15 minutes.
• If using a 4mL stock microsphere vial, rotate the vial for 15 minutes at 15-30
rpm.