Bio-Rad Chromatographic Surfaces - Manual (Page 3)

Technical Considerations
I Prerinsing the ProteinChip NP20 array with HPLC or better
grade water may improve results
Protocol: On-Spot
Note: This protocol is intended as a guideline; you may need to optimize the method
for your particular sample type and experimental design.
1. Dilute or dissolve the sample in binding buffer. Most buffers are
tolerated by this surface.
2. Load 5 µl of sample per spot (ideally 200–2,000 nM, 10-fold less
for peptides).
3. Allow sample to air-dry onto the surface for 5–10 minutes.
4. Wash each spot with 5 µl of binding buffer with shaking
(on MicroMix shaker, setting 20/4) and remove buffer.
Repeat two more times.
5. Air-dry the array for 15–20 minutes.
6. Apply 0.5 µl of saturated ProteinChip energy absorbing
molecules (EAM) in solution to each spot. Allow to air-dry.
Optional: Add another 0.5 µl of saturated EAM in solution to
improve signal intensity.
7. Analyze the arrays using the ProteinChip SELDI system.
© 2006 Bio-Rad Laboratories, Inc.