Bio-Rad Chromatographic Surfaces - Manual (Page 6)

© 2006 Bio-Rad Laboratories, Inc.
Protocol 2: Serum Profiling On-Spot
1. Optional: Bulk-wash the ProteinChip array with 50% methanol or
acetonitrile for five minutes. Repeat once. Dry the array for an
hour after bulk wash to minimize any spot-to-spot
cross-contamination.
2. Prewet the spots with 5 µl of binding buffer for 2 minutes.
Repeat once.
3. Remove the prewetting solution and replace with 5 µl of sample.
Do not allow the spot to air-dry during sample application.
4. Incubate in a humid chamber for 30 minutes with shaking
(on MicroMix shaker setting 20/4).
5. Wash each spot with 5 µl binding buffer for 2 minutes with
shaking, and remove buffer. Repeat two more times.
6. Optional: If binding buffer contains salt, wash each spot with 5 µl
of DI water.
7. Air-dry the array for 5–10 minutes.
8. Apply 1 µl of ProteinChip EAM in solution to each spot. Air-dry
for 5 minutes, and apply another 1 µl of EAM in solution. Allow
to air-dry.
9. Analyze the array using the ProteinChip SELDI system.