Fig. 6: Feasibility of f-NTA for the detection of physiologically relevant proteins. | Communications Biology

Fig. 6: Feasibility of f-NTA for the detection of physiologically relevant proteins.

From: Tracking matricellular protein SPARC in extracellular vesicles as a non-destructive method to evaluate lipid-based antifibrotic treatments

Fig. 6

a SEC-purified EVs stained with unspecific membrane dye PKH67 by systematically increasing dye concentrations. b SEC-purified EVs incubation with varying amounts of AF488-CD81 and for different times at 24 °C. c, d EV-containing pellets incubated with AF488-CD81 prior to SEC and detected after purification (c) and comparison with additional incubation with AF488-CD9 (d). e Direct comparison of CD9 detection by f-NTA and FACS using EVs from differently treated LX-2, including isotype control (IC). f Detection of SPARC and CD81 on EVs isolated from differently treated LX-2 cells following the optimized f-NTA protocol (mean ± SD, n = 3). P values (p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), and p ≤ 0.0001 (****)) were determined by one-way ANOVA on ranks and Tukey’s multiple comparison.

Back to article page