Bio-Rad Matrix-Prepared Surfaces - Manual (Page 6)

5 © 2007 Bio-Rad Laboratories, Inc.
Protocol 1
The following method can be used to apply a sample directly to
the ProteinChip SEND ID array for peptide analysis.
1. Mix your sample 1:1 (v/v) with 50% ACN (v/v) and 0.2% TFA
in deionized (DI) water.
2. Add 1–2 µl of sample to each spot on the ProteinChip
SEND ID array.
3. Allow the spots to air-dry.
4. Read the arrays in the ProteinChip SELDI reader.
Protocol 2
The following method can be used to clean up a sample on
the ProteinChip array to remove salt and urea contamination
(see Figure 4). The C-18 chemistry on the surface selectively retains
peptides through their hydrophobic regions while impurities are
washed away.
1. Mix sample with 0.2% TFA at a 1:1 (v/v) ratio.
2. Apply 5 µl of 0.1% TFA to the ProteinChip SEND ID array.
Remove within 30 sec; do not agitate. Repeat.
3. Spot 2 µl of sample onto the array.
4. Incubate the samples for 10 min in a humidity chamber
at room temperature. Remove the samples.
5. Add 5 µl of 0.1% TFA to each spot. Remove within
30 sec; do not agitate.
6. Add 2 µl of 25% ACN (v/v) and 0.1% TFA in DI water to
each spot.
7. Allow the array to air-dry.
8. Read the array in the ProteinChip SELDI reader.