Table 17.
Washing Twice with Buffer Containing 50 mM ABC
| Processing time | Step | Comments / Tips |
|---|---|---|
| 1–2 hours | Add 200 μL 50 mM ABC (only ABC) to each sample filtration unit. Centrifuge units for 15 minutes at 12,000 × g. | Reconstitute trypsin during last wash step. Particularly for the last wash step, increase the centrifugation speed to 12,500 × g or 13,000 × g if need be to pass all the wash solution through the filter. |
| Empty eluent into waste for the second time so that it does not reach a volume that touches the filter. Handle the filter very carefully. | ||
| Repeat the wash with another 200 μL 50 mM ABC (only) buffer. Centrifuge units for 15 minutes at 12,000 × g. |