Luminex 100 User Manual Version 1.7 - Manual (Page 115)

x MAP Technology Protocols
PN 89-00002-00-063 Rev. A 9 - 5
Two-Step
Carbodiimide
Coupling of Protein to
xMAP Carboxylated
Microspheres
Introduction
Use the protocols included here as a general starting point for
developing assays. All assays should be optimized for your reagents
in your specific application. You will discover the methods that give
you the best results by starting with these guidelines and modifying
them for your specific needs.
Equipment
• Vortex
• Sonicator bath
• Micropipetters (1 µl - 1000 µl)
• Microcentrifuge
• Analytical balance
• Timer
• Rotator
Materials
• xMAP carboxylated microspheres—LIMIT EXPOSURE TO
LIGHT!
• Microcentrifuge tubes: 1.5 ml, polypropylene (See Technical
note 4)
• ACTIVATION BUFFER:
0.1 M Monobasic Sodium Phosphate (pH 6.2 ± 0.2)
• COUPLING BUFFER:
50 mM MES (pH 5.0 - 6.0)
• WASH BUFFER:
Phosphate Buffered Saline (pH 7.4 ± 0.1), Tween
®
20
(0.05% v/v)
• BLOCKING/STORAGE BUFFER:
Phosphate Buffered Saline (pH 7.4 ± 0.1), Bovine Serum
Albumin (10 mg/mL), Sodium Azide (0.05% w/v)
• Sulfo-NHS:
N-Hydroxysulfosuccinimide sodium salt, Pierce Chemicals
• EDC:
1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride,
Pierce Chemicals
• Protein for coupling (See Technical note 2)