Luminex 100 IS Developer Workbench Guide Version 2.3 - Manual (Page 65)

PN 89-00002-00-084 Rev. B 59
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MAP Technology LumAvidin Binding Protocol
LumAvidin Binding 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
• Micropipetters (1 µL - 1000 µL)
• Microcentrifuge
• Timer
• Rotator
Materials
• xMAP LumAvidin-modified xMAP microspheres—LIMIT
EXPOSURE TO LIGHT!
• Biotin-conjugated molecule
• Microcentrifuge tubes: 1.5 mL, polypropylene (see technical
note 3)
• REACTION BUFFER:
Phosphate Buffered Saline (pH 7.3 ± 1%), Bovine Serum
Albumin (BSA)
• 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 xMAP microsphere stock for 1
minute at
≥ 8,000 × g.
2. Disperse the pellet with sonication, and vortex the container for
20 seconds.
3. Dispense 1.0
× 10 5 Avidin-modified xMAP microspheres into a
1.5 mL USA Scientific microcentrifuge tube.
4. Centrifuge the xMAP microspheres for 1 minute at
≥ 8,000 × g.
™
Note: We recommend titration
in the 4 to 4000 nM range to
determine the optimal amount of
biotin-conjugated molecule per
specific binding reaction.