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. Author manuscript; available in PMC: 2023 Mar 6.
Published in final edited form as: Rapid Commun Mass Spectrom. 2021 Dec 23;38(Suppl 1):e9189. doi: 10.1002/rcm.9189

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.