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Membrane markers for EV and MFGM. (A) EVs and MFGM were assessed for the presence of six selected, well‐established membrane markers by western blotting. It is observed that CD9, CD63 and CD81 are more strongly detected in the EV sample, with the opposite being observed for BTN, XOD and LAH. (B) Representative immunostain electron microscopy pictures of BTN and CD9 binding to a wide range of samples. All scalebars are 200 nm except for the WFC samples, where it is 500 nm. The black spots are the gold particles bound to the secondary antibodies, while the ‘shadows’ are sample material. An overview of the samples and accompanying labels can be found in Figure S2.

Journal: Journal of Extracellular Biology

Article Title: Industrial Scale Production and Characterization of a Whey Fraction Enriched in Extracellular Vesicle Material

doi: 10.1002/jex2.70044

Figure Lengend Snippet: Membrane markers for EV and MFGM. (A) EVs and MFGM were assessed for the presence of six selected, well‐established membrane markers by western blotting. It is observed that CD9, CD63 and CD81 are more strongly detected in the EV sample, with the opposite being observed for BTN, XOD and LAH. (B) Representative immunostain electron microscopy pictures of BTN and CD9 binding to a wide range of samples. All scalebars are 200 nm except for the WFC samples, where it is 500 nm. The black spots are the gold particles bound to the secondary antibodies, while the ‘shadows’ are sample material. An overview of the samples and accompanying labels can be found in Figure S2.

Article Snippet: The following antibodies were used: monoclonal mouse anti‐human CD9 (clone IVA50, reducing conditions; diluted 1:1000; ThermoFischer), monoclonal mouse anti‐bovine CD63 (clone CC25; non‐reducing conditions; diluted 1:2000; ThermoFischer), monoclonal mouse anti‐human CD81 (clone 12C4; non‐reducing conditions; diluted 1:1000; CosmoBio), monoclonal rabbit anti‐bovine BTN (clone 102.43; reducing conditions; diluted 1:1000; LSBio), polyclonal rabbit anti‐human XOD (reducing conditions; diluted 1:1000, Abcam), and polyclonal rabbit anti‐bovine lactadherin (reducing conditions; diluted to 1 μg/mL; in house‐produced by Jan Trige Rasmussen, Aarhus University).

Techniques: Membrane, Western Blot, Electron Microscopy, Binding Assay

Overview of the peptides used for quantification by mass spectrometry.

Journal: Journal of Extracellular Biology

Article Title: Industrial Scale Production and Characterization of a Whey Fraction Enriched in Extracellular Vesicle Material

doi: 10.1002/jex2.70044

Figure Lengend Snippet: Overview of the peptides used for quantification by mass spectrometry.

Article Snippet: The following antibodies were used: monoclonal mouse anti‐human CD9 (clone IVA50, reducing conditions; diluted 1:1000; ThermoFischer), monoclonal mouse anti‐bovine CD63 (clone CC25; non‐reducing conditions; diluted 1:2000; ThermoFischer), monoclonal mouse anti‐human CD81 (clone 12C4; non‐reducing conditions; diluted 1:1000; CosmoBio), monoclonal rabbit anti‐bovine BTN (clone 102.43; reducing conditions; diluted 1:1000; LSBio), polyclonal rabbit anti‐human XOD (reducing conditions; diluted 1:1000, Abcam), and polyclonal rabbit anti‐bovine lactadherin (reducing conditions; diluted to 1 μg/mL; in house‐produced by Jan Trige Rasmussen, Aarhus University).

Techniques: Sequencing

MFGM and EV marker proteins used in this study.

Journal: Journal of Extracellular Biology

Article Title: Industrial Scale Production and Characterization of a Whey Fraction Enriched in Extracellular Vesicle Material

doi: 10.1002/jex2.70044

Figure Lengend Snippet: MFGM and EV marker proteins used in this study.

Article Snippet: The following antibodies were used: monoclonal mouse anti‐human CD9 (clone IVA50, reducing conditions; diluted 1:1000; ThermoFischer), monoclonal mouse anti‐bovine CD63 (clone CC25; non‐reducing conditions; diluted 1:2000; ThermoFischer), monoclonal mouse anti‐human CD81 (clone 12C4; non‐reducing conditions; diluted 1:1000; CosmoBio), monoclonal rabbit anti‐bovine BTN (clone 102.43; reducing conditions; diluted 1:1000; LSBio), polyclonal rabbit anti‐human XOD (reducing conditions; diluted 1:1000, Abcam), and polyclonal rabbit anti‐bovine lactadherin (reducing conditions; diluted to 1 μg/mL; in house‐produced by Jan Trige Rasmussen, Aarhus University).

Techniques: Marker, Membrane