Journal: bioRxiv
Article Title: Combining Anion Exchange and Size Exclusion Chromatography for Extracellular Vesicle Enrichment from Small Volumes of Human and Mouse Plasma for Quantitative Proteomics
doi: 10.64898/2026.03.11.711200
Figure Lengend Snippet: SEC Void optimisation. Pooled SEC fractions 1-4 collected using a 2.7 mL void volume (1-4A) and a 2.8 mL void volume (1-4B) were prepared in triplicate from 0.5 mL plasma. Western blot analysis of contaminant proteins Albumin (green) and lipoprotein ApoA1 (red) (A) with 0.5 µl plasma included as a positive control (+). EV markers CD63 and syntenin-1 and small EV exclusion marker Calnexin (B). Samples (0.5 ml and 0.4 ml) from Experiment 3.2 included as a western blot control and 2µl platelet lysate included as a calnexin positive control (+). Electron microscopy (C) visualisation and size and concentration histogram (D) of EV representative SEC sample (Void 2.7 ml).
Article Snippet: Membranes were blocked in 5% skim milk in PBS at room temperature for 1 hour before overnight incubation with primary antibodies Syntenin (Abcam, cat# ab133267), CD63 (Abcam, cat#ab134045), CD63 (Abcam, cat#ab8219), Albumin (Cell Signalling Technology, cat#4929S), Calnexin (Cell Signalling Technology, cat#2679S) ApoA1 (Santa Cruz, cat# SC-376818).
Techniques: Clinical Proteomics, Western Blot, Positive Control, Marker, Control, Electron Microscopy, Concentration Assay