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However, re\u2010solubilization of the total protein precipitate is difficult using regular in\u2010solution digestion protocol. Sodium deoxycholate is reported as an efficient surfactant for re\u2010solubilization of membrane fractions. In this study, we demonstrated an application combining methanol\u2013chloroform based protein precipitations and deoxycholic acid assisted re\u2010solubilization of pellets to evaluate the improvement of protein identifications in mass spectrometry\u2010based bottom\u2010up proteomics. We evaluated the modified method using an equal amount of Raw 264.7 mouse macrophage cell lysate. Detailed in\u2010solution trypsin digestion studies were presented on methanol\u2013chloroform precipitated samples with or without deoxycholic acid treatments and compared with popular sample digestion methods. A mass spectrometric analysis confirmed an 82% increase in protein identification in deoxycholic acid\u2010treated samples compared to other established methods. Furthermore, liquid chromatography\u2013tandem mass spectrometry analysis of an equal amount of proteins from methanol\u2013chloroform precipitated, and methanol\u2013chloroform\/deoxycholic acid\u2010treated macrophage cell lysate showed a 14% increase and 27% unique protein identifications. We believe this improved digestion method could be a complementary or alternative method for mammalian cell sample preparations where sodium dodecyl sulfate based lysis buffer is frequently used.<\/jats:p>","DOI":"10.1002\/jssc.201901273","type":"journal-article","created":{"date-parts":[[2020,2,19]],"date-time":"2020-02-19T14:42:01Z","timestamp":1582123321000},"page":"2125-2132","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Improved in\u2010solution trypsin digestion method for methanol\u2013chloroform precipitated cellular proteomics sample"],"prefix":"10.1002","volume":"43","author":[{"given":"A. D. 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