Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Extracellular Vesicle Marker Changes Associated With Disease Activity in Relapsing-Remitting Multiple Sclerosis

doi: 10.1212/NXI.0000000000200404

Figure Lengend Snippet: EVs were isolated by size-exclusion chromatography (SEC, SmartSEC, System Biosciences). (A) Analysis of the classical EV marker Alix by Western blotting after isolation. Equal amounts of EVs and plasma samples (10 µg) were used. A weak level of albumin was detected in the EV fraction. (B) Freshly isolated EVs were subjected to cryo-electron microscopy (cryo-EM). Cryo-electron microscopy observations showed EVs with typical lipid bilayer membrane. EV = extracellular vesicle.

Article Snippet: Thereafter, membranes were incubated with a primary antibody against Alix (1:500, NBP1-90201, Novus Biologicals, Centennial, CO) in a rolling shaker overnight at 4°C.

Techniques: Isolation, Size-exclusion Chromatography, Marker, Western Blot, Clinical Proteomics, Cryo-Electron Microscopy, Cryo-EM Sample Prep, Membrane

Journal: Future microbiology

Article Title: Liquid biopsy in the diagnosis of HPV DNA in breast lesions.

doi: 10.2217/fmb-2017-0145

Figure Lengend Snippet: Figure 1. Serum-derived extracellular vesicles isolation and characterization. (A) Serum-derived EVs were isolated by differential ultracentrifugation and DNase-I digested, (B) Western Blot and NanoSight analysis of 8 Alix positive serum-derived EVs (see Table 2) and representative NanoSight plot (n = 3 replicates) of sample 51 (see Table 2). EV: Extracellular vesicle.

Article Snippet: The effective presence of EVs was assessed by Western Blot for Alix expression (Mouse antiALIX antibody, clone 3A9, MCA2493, Bio-Rad, Milan Italy) and EVs were counted by NanoSight NS500 (IRST, Meldola, Italy) (Figure 1B).

Techniques: Derivative Assay, Isolation, Western Blot

Journal: iScience

Article Title: An antibiotic derivative as a new potential tool in the prevention of hemolytic uremic syndrome

doi: 10.1016/j.isci.2025.113076

Figure Lengend Snippet: Quantitative capillary western blot analysis of the proteins derived from human EVs Human EVs were isolated after treatment of human blood with vehicle or with 2 nM Stx2a in the absence or in the presence of 0.01 μg/mL NAB815 and their proteins extracted as described in . (A) Representative WES of the analyzed antigens (Alix, CD45, and CD42a) and associated proteins (Stx2a) is shown. (B–E) Data obtained with different human donors ( n = 4) represent the percentage (mean ± SD) of the quantitative determinations of the different antigens with respect to controls (C–E) or Stx2a (B). (F–H) Data obtained with different human donors ( n = 4) are expressed as percentage (mean ± SD) of the stimulation in the presence of Stx2a that was set at 100%. Different primary antibodies were used: rabbit polyclonal anti-Alix 1:50 (Novus Biological) as an EV marker, mouse monoclonal anti-CD45 1:250 (BD Transduction Laboratories) as a leukocyte marker, rabbit polyclonal anti-CD42a 1:10 (GeneTex) as a platelet marker, and rabbit polyclonal anti-Stx2a 1:50 (Dr. Stefano Morabito, ISS Rome) to detect the toxin. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 (two-tailed unpaired t test). See also .

Article Snippet: Different primary antibodies were used: rabbit polyclonal anti-Alix 1:50 (Novus Biological) as an EV marker, mouse monoclonal anti-CD45 1:250 (BD Transduction Laboratories) as a leukocyte marker, rabbit polyclonal anti-CD42a 1:10 (GeneTex) as a platelet marker, and rabbit polyclonal anti-Stx2a 1:50 (Dr. Stefano Morabito, ISS Rome) to detect the toxin.

Techniques: Western Blot, Derivative Assay, Isolation, Marker, Transduction, Two Tailed Test