Luminex 100 IS Version 2.2 - Manual (Page 90)

Luminex 100 IS Developer Guide to x MAP Technology Version 2.2 x MAP Technology
84 PN 89-00002-00-032 Rev. A
Binding Biotin-conjugated Molecules to 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
• x MAP 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
1. 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 10,000 × g.
2. Disperse the pellet with sonication, and vortex the container for
20 seconds.
3. Dispense 1.0 × 10
5
Avidin-modified microspheres into a 1.5 mL
microcentrifuge tube.
4. Centrifuge the microspheres for 1 minute at 10,000 × g and
remove the supernatant.