Source: Collins, S. M., et al., Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity. J. Vis. Exp. (2021).
In this video, we demonstrate a technique–size exclusion chromatography–to separate a heterogenous population of outer membrane vesicles or OMVs derived from bacteria. These vesicles are nanosized spherical structures and are separated into large and small subpopulations based on their size through Size Exclusion Chromatography.
1. Preparation of buffers
2. Preparation of OMV sample
3. Packing the S-1000 column
4. Loading the sample and collecting fractions
The authors have nothing to disclose.
Amicon 50 kDa filters | Millipore Sigma | UFC905024 | |
FM 4-64 | Thermo Scientific | T13320 | 1.5 x 50 cm |
Glass Econo-Column | BioRad | 7371552 | |
Potassium Chloride (KCl) | Amresco (VWR) | 0395-500G | |
Potassium Phosphate Monobasic Anhydrous (KH2PO4) | Amresco (VWR) | 0781-500G | |
Sephacryl S-1000 Superfine | GE Healthcare | 17-0476-01 | |
Sodium Chloride (NaCl) | Fisher Chemical | S271-3 | |
Sodium Phosphate Dibasic Anhydrous (Na2HPO4) | Amresco (VWR) | 0404-500G | |
Tris Base | VWR | 0497-1KG |