iodixanol solution optiprep tm millipore d1556  (Millipore)

Journal: bioRxiv

Article Title: Isolating Small Extracellular Vesicles from Small Volumes of Blood Plasma using size exclusion chromatography and density gradient ultracentrifugation: A Comparative Study

doi: 10.1101/2023.10.30.564707

Figure Lengend Snippet: (A) SEC-PFs (6 ml ) were placed on top of a density gradient cushion constructed with 2 ml 10%, 2 ml 30%, and 2 ml 50% iodixanol solutions and centrifuged at 150,000 × g for 2 h at 4 °C . (B) After DGUC, the tube was fractionated into 42 fractions, which were each examined for their densities, particle concentrations (by NTA) and presence of sEVs and lipoproteins (by TEM). The dominance of lipoproteins was evident in fractions of density <1.05 g/ml, which is designated as LP zone. sEVs started to appear when density exceeded 1.05 g/ml but a significant level of lipoproteins was present until the density of 1.08 g/ml . Therefore, the density region of 1.05~1.08 g/ml was designated as LP+sEV zone. Beyond 1.08 g/ml , lipoproteins diminished drastically and the density region of >1.08 g/ml was designated as the sEV zone. All scale bars represent 200 nm .

Article Snippet: To prepare the density solutions, a 50% Working Solution was firstly made by diluting 5 vol of iodixanol (Optiprep TM , D1556, Sigma Aldrich, MO, USA) with 1 vol of 0.25 M Sucrose, 6 mM EDTA, 60 mM Tris-HCl (pH 7.4).

Techniques: Construct

Journal: bioRxiv

Article Title: Isolating Small Extracellular Vesicles from Small Volumes of Blood Plasma using size exclusion chromatography and density gradient ultracentrifugation: A Comparative Study

doi: 10.1101/2023.10.30.564707

Figure Lengend Snippet: (A) 500 μl PBS was overlaid on 800 μl 10% iodixanol solution and 20 μl 50% iodixanol cushion. The 1.5 ml tubes were subjected to ultracentrifugation using a fixed-angle rotor at an average speed of 135,000 g for 2, 6 and 16 h at 4 °C to establish a density gradient profile. (B) Density gradient profiles along the 1.5 ml tubes after ultracentrifugation. The 2 h spinning time gave a density profile with the largest separation zone of 1.05~1.08 g/ml and smallest sEV zone of >1.08 g/ml . The data shows the average ± standard deviation of 7 repeats for 2 h , 3 repeats each for 6 and 16 h .

Article Snippet: To prepare the density solutions, a 50% Working Solution was firstly made by diluting 5 vol of iodixanol (Optiprep TM , D1556, Sigma Aldrich, MO, USA) with 1 vol of 0.25 M Sucrose, 6 mM EDTA, 60 mM Tris-HCl (pH 7.4).

Techniques: Standard Deviation