Luminex 100 IS Version 2.1 User Manual
Page 80
Luminex 100 IS Developer Guide to xMAP Technology Version 2.1
x
MAP
Technology
74
PN 89-00002-00-062 Rev. A
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
≥ 8,000 × g 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
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.
Aspirate the supernatant.
9. Add 500 µL of BLOCKING/STORAGE BUFFER to the
microspheres and vortex. Centrifuge the tube for 60 seconds.
Aspirate the supernatant.
10. Repeat step 9.
11. Resuspend the microspheres in the desired volume of
BLOCKING/STORAGE BUFFER.
12. Enumerate the microsphere preparation.
13. Store the preparation at 2
o
C - 8°C. Protect it from light.
Technical Notes
1. The concentration of Biotin-conjugated molecules is provided as
a guide for assay development. The concentration required for
optimal reactivity must be determined empirically for each
specific biotin-conjugated molecule.
2. The reaction volume can vary with the reagent volume. If the
reagent is limited, perform the reaction in a proportionately low
volume.