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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 11.

Centrifugation and Removal of Lysate

Processing time Step Comments / Tips
2–3 hours Centrifuge all samples at 16,000 × g for 30 minutes at 4°C to form pellets. With high quantities of tissue in 10 mL lysis buffer, concentration risks being higher than the calibration curve of the Pierce BCA Protein Assay Kit. It is therefore recommended to create a 1:10 diluted sample in Lysis Buffer prior to the total protein quantification assay analysis.
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Remove lysate to a fresh tube and label. Care should be taken not to disturb the pellet. Lysates should be kept on ice, and pellets can be stored at −80°C for potential future analysis, if needed. Lysates of samples are now ready for protein quantitation and digestion.
Quantitation of protein can be performed using the Pierce BCA Protein Assay Kit or using the Nanodrop A280 protein assay. Lysis buffer should be used as a blank. Following protein quantitation, lysates should be stored at −80°C until the beginning of the digestion procedure.