Luminex 100 IS Developer Workbench Guide Version 2.3 - Manual (Page 58)

Luminex 100 IS Developer Workbench Guide Version 2.3
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MAP Technology
52 PN 89-00002-00-084 Rev. B
Protein Coupling Protocol
Introduction
Use these protocols as a general starting point for developing assays.
Optimize all assays for your reagents in your specific application.
Updates or additions to these protocols are posted on the Luminex
website at
http://luminexcorp.custhelp.com. At the main page, select
a subject or perform a search for the desired information.
You obtain 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 1)
• Activation Buffer:
0.1 M Sodium Phosphate (pH 6.2 ± 0.2)
• Coupling Buffer:
50 mM MES (pH 5.0) (see Technical note 2)
• 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 (see
Technical note 3)
• EDC:
1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride,
Pierce Chemicals (see Technical note 3)
• Protein for coupling (see Technical note 4)