Luminex 100 IS Version 2.1 - Manual (Page 79)

PN 89-00002-00-062 Rev. A 73
x MAP Technology
3. The optimal coupling concentration for a given oligonucleotide
is determined by coupling at various concentrations within the
recommended range of 1 to 50 µM.
4. Minimize the exposure of EDC to air; secure closures on stock
and dry aliquot containers, store desiccated at -20°C. Use
aliquots immediately and discard containers after use. Make a
fresh 10 mg/mL EDC solution before each addition.
Binding Biotin-conjugated Molecules to xMAP
LumAvidin-Modified 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
• Micropipetters (1 µL - 1000 µL)
• Microcentrifuge
• Timer
• Rotator
Materials
• xMAP LumAvidin-modified microspheres—LIMIT
EXPOSURE TO LIGHT!
• Biotin-conjugated molecule
• Microcentrifuge tubes: 1.5 mL, polypropylene
• REACTION BUFFER:
Phosphate Buffered Saline (pH 7.3 ± 0.1), Bovine Serum
Albumin (10 mg/mL)
• BLOCKING/STORAGE BUFFER:
Phosphate Buffered Saline (pH 7.3 ± 0.1), Tween 20
(0.02% v/v), Bovine Serum Albumin (1 mg/mL), Sodium Azide
(0.05% w/v)
Preparation
Dilute the Biotin-conjugated molecule in REACTION BUFFER to a
concentration of 4 - 4000 nM (See Technical note 1).
Procedure
1. Centrifuge the Avidin-modified microsphere stock for 1 minute
at ≥ 8,000 × g.