Bio-Rad Chromatographic Surfaces User Manual

Page 5

Protocol 1: Serum Profiling Using the Bioprocessor

Note: These protocols are intended as a guideline; you may need to optimize the
method for your particular sample type and experimental design.

Note: This protocol is for the 8-spot array in the ProteinChip cassette-compatible
bioprocessor. For processing a single array, use the ProteinChip 8-well bioprocessor
(catalog #C50-30008).

1. Place the ProteinChip array cassette in the bioprocessor, and

prewash the arrays by adding 50 µl 50% methanol or
acetonitrile for five minutes. Repeat once.

2. Remove the prewash solution from the wells and add

150–250 µl binding solution to each well. Incubate for
5 minutes at room temperature with vigorous shaking (e.g.,
250 rpm, or on MicroMix shaker setting 20/7). Repeat once.

3. Remove the buffer from the wells. Immediately add 50–150 µl

sample to each well. Recommended concentration is
50–2,000 µg/ml total protein, in binding buffer. Incubate with
vigorous shaking for 30 minutes.

4. Remove the samples from the wells and wash each well with

150–250 µl binding buffer for 5 minutes, with agitation. Repeat
two more times.

5. Remove the binding buffer from the wells and add 150–250 µl

deionized (DI) water to each well. Remove immediately.

6. Remove the reservoir from the bioprocessor base clamp

assembly.

7. Air-dry the arrays for 5–10 minutes.

8. Add ProteinChip energy absorbing molecules (EAM) after

removing the reservoir; use the cassette hold-down frame
provided with the ProteinChip cassette-compatible
bioprocessor to keep the cassette flat during EAM addition.

9. 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.

10. Analyze the arrays using the ProteinChip SELDI system.