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. 2017 Mar 15;6(1):1294340. doi: 10.1080/20013078.2017.1294340

Figure 2.

Figure 2.

SEC elution profiles and EV markers in human and bovine fluff and milk serum fractions. SEC (size-exclusion chromatography) profiles of human (a) and bovine (b) fractions were obtained by recording the absorbance at 280 nm. Fractions across the SEC profiles were collected from the fluff (F, in red) and milk serum (S, in green) runs with the eluting peaks annotated with F1–F4 and S1–S5. The solid casein rich pellets derived from the 340,000 g ultracentrifugation (340K pellet) and the 20,000 g centrifugation (20K pellet) were collected and included in the analysis as well as an MFGM preparation. All samples were analysed for their content of lactadherin, CD9, CD63 and CD81 by Western blotting and/or LC-MS. Moreover, human milk SEC fractions were screened for β-casein content by Western blotting, and bovine samples for their content of β-casein, αS1- and αS2-casein by LC-MS. Ten µg protein from the indicated SEC fractions were analysed by Western blotting using antibodies targeting the indicated proteins with their approximated weights shown. Relative levels obtained by LC-MS are based on the detection of peptides specifically derived from the indicated proteins. Abbreviations: MFGM, milk fat globule membrane; LC-MS, liquid chromatography–mass spectrometry.