Materials, Preparation, Procedure – Luminex 200 User Manual with LDS 1.7 Software User Manual

Page 128

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Luminex 200 User Manual for LDS Version 1.7

x

MAP Technology

9 - 10

PN 89-00002-00-150 Rev. A

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

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 xMAP microsphere stock for 1

minute at 8,000

× g or higher.

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 microcentrifuge tube.

4. Centrifuge the xMAP microspheres for 1 minute at 8,000

× g or

higher and remove the supernatant.

5. Add 250 µl of the REACTION BUFFER (See Technical

note 2). Vortex.

6. Add 250 µl of the Biotin-conjugated molecule solution to the

xMAP microsphere suspension. Vortex immediately.

7. Rotate the mixture for 30 minutes in the dark at room

temperature.

8. Centrifuge the microspheres for 1 minute at 8,000

× g or higher.

Aspirate the supernatant.

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