Journal: bioRxiv

Article Title: Standardized brain and plasma EV enrichment pipeline validated for Single sample multi-Omic and fatty acids applications in Mouse and Human

doi: 10.64898/2026.01.22.700328

Figure Lengend Snippet: (A-C) Comparison of average particle concentration, size and free protein concentration from fractions III-IX between collagenase and papain treated mouse brains. (D-F) Comparison of the percentage of CD81 + , CD9 + and CD63 + events in fractions III-IX between collagenase and papain treated mouse brains. (G-H) Comparison between the percentage of CD81 + , CD9 + and CD63 + events in fractions III-IX from mouse and human (I-J) Free protein concentration (ug/mL) in fractions I-IV of plasma EV isolates in mouse and human. Data are expressed as mean ± SEM, collagenase n=4, papain n=4, Two-way ANOVA followed by Bonferroni’s post-hoc analysis. Statistical differences: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The following primary antibodies were used: CD9 (Merck, CBL162); CD63 (BioOrbyt, orb229829); TSG101 (Santa Cruz Biotechnology, sc-7964); β-Actin (BioLegend, 622102); Cytochrome C (Cell Signaling Technology/Ozyme, 11940); Histone H2A.Z (Cell Signaling Technology/Ozyme, #2718); CD81 (Cell Signaling Technology/Ozyme, 10037); CD81 (Cell Signaling Technology/Ozyme, 56039); Alix (Cell Signaling Technology/Ozyme, #2717S); CD9 (Cell Signaling Technology/Ozyme, 98327); CD63 (Ozyme, CAB5271); L1CAM (Abcam, ab24345); APOE (Cell Signaling Technology, #13366).

Techniques: Comparison, Concentration Assay, Protein Concentration, Clinical Proteomics

Journal: bioRxiv

Article Title: Standardized brain and plasma EV enrichment pipeline validated for Single sample multi-Omic and fatty acids applications in Mouse and Human

doi: 10.64898/2026.01.22.700328

Figure Lengend Snippet: (A) Representation of BDEV and PDEV from human and mouse by TEM. (B) Size distribution of BDEVs by TEM from human and mouse. (C) Representative size distribution of BDEVs and PDEVs from human and mouse by NanoFCM. (D-E) Comparison of mean concentration and size of BDEV and PDEVs (MBDEV=4, MPDEV=13, HBDEV=4, HPDEV=6). (F) Representative dot plots displaying CD81+, CD9+ and CD63+ events from HBDEVs. Analysis of CD81, CD9 and CD63 expression (%) in BDEVs and PDEVs by NanoFCM (n=4). (G-H) Representative immunoblots of commonly detected proteins in EVs (CD81, CD9, CD63, TSG101 and L1CAM) and negative control proteins (Histone H2A, Cytochrome C and APOE) in BDEV and PDEVs from human and mouse (H=Homogenate, H+C=Homogenate + Collagenase IV, P=Plasma) . Data are expressed as mean ± SEM, One-way ANOVA followed by Bonferroni’s post-hoc analysis for multiple (F) and pairwise comparisons (D and E). Statistical differences: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Scale bar is 200 nm.

Article Snippet: The following primary antibodies were used: CD9 (Merck, CBL162); CD63 (BioOrbyt, orb229829); TSG101 (Santa Cruz Biotechnology, sc-7964); β-Actin (BioLegend, 622102); Cytochrome C (Cell Signaling Technology/Ozyme, 11940); Histone H2A.Z (Cell Signaling Technology/Ozyme, #2718); CD81 (Cell Signaling Technology/Ozyme, 10037); CD81 (Cell Signaling Technology/Ozyme, 56039); Alix (Cell Signaling Technology/Ozyme, #2717S); CD9 (Cell Signaling Technology/Ozyme, 98327); CD63 (Ozyme, CAB5271); L1CAM (Abcam, ab24345); APOE (Cell Signaling Technology, #13366).

Techniques: Comparison, Concentration Assay, Expressing, Western Blot, Negative Control, Clinical Proteomics

Journal: eLife

Article Title: Framework for rapid comparison of extracellular vesicle isolation methods

doi: 10.7554/eLife.70725

Figure Lengend Snippet:

Article Snippet: The following recombinant proteins were used for CD9, CD63, CD81, and albumin: ab152262 (Abcam), TP301733 (Origene), TP317508 (Origene), and ab201876 (Abcam).

Techniques: Clinical Proteomics, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Recombinant, Isolation

Journal: Blood Advances

Article Title: Polyphosphate expression by cancer cell extracellular vesicles mediates binding of factor XII and contact activation

doi: 10.1182/bloodadvances.2021005116

Figure Lengend Snippet: Characterization of EV. Particle numbers (A) and relative protein concentrations (B) in the fractions eluted following qEV size-exclusion chromatography. Data are mean ± SEM. (C) Tetraspanin (CD9, CD63, and CD81) content of purified EVs was analyzed in the indicated elution fractions by immunoblotting. (D) Size distribution of pooled fractions 7 to 10 from HDFs (left panel) and L3.6 cells (right panel). (E) Electron microscopy image of EV from HDFs (left panel) and L3.6 cells (right panel).

Article Snippet: Anti-human CD81-AF488 antibody (20 μg/mL, FAB4615G; R&D Systems) was used as a control ligand.

Techniques: Size-exclusion Chromatography, Purification, Western Blot, Electron Microscopy

Journal: Blood Advances

Article Title: Polyphosphate expression by cancer cell extracellular vesicles mediates binding of factor XII and contact activation

doi: 10.1182/bloodadvances.2021005116

Figure Lengend Snippet: polyP on EV derived from cancer cells mediates ligand binding. (A) The amount of polyP associated with cancer cell–derived EV was estimated by comparison with a standard curve (supplemental Figure 2A). (B) Detection of polyP on L3.6-derived EVs following incubation with buffer (HBS), DNase I and RNase A (D/R), or CIP. polyP of different chain lengths was used to estimate the size of EV-associated polyP. L3.6-derived EVs were preincubated with buffer or E coli PPX, and the binding of PPX-Δ12–AF488 (C), anti-CD81–AF488 (D), and or FXII-AF488 (E) was measured. The number of total EV was determined in scatter mode, and the number of ligand-binding EV was determined in fluorescence mode using a 488-nm excitation laser. The differences in fluorescent particle numbers were compared by estimating the area under each curve (AUC) using GraphPad Prism 8. Bars represent means ± SEM. *** P < .001, 1-way ANOVA with multiple comparisons. L, long; M, medium; S, short.

Article Snippet: Anti-human CD81-AF488 antibody (20 μg/mL, FAB4615G; R&D Systems) was used as a control ligand.

Techniques: Derivative Assay, Ligand Binding Assay, Comparison, Incubation, Binding Assay, Fluorescence