Magnetic Bead-based Exosome Extraction: A Technique to Isolate Specific Exosomes from Biofluids
Published: April 30, 2023
Abstract
Source: Tu, M. et al. Detection of Exosomal Biomarkers by Electric Field-induced Release and Measurement (EFIRM). J. Vis. Exp. (2015)
In this video, we demonstrate a procedure to isolate exosomes from target biofluids using magnetic microbeads. The procedure enables the rapid capture of exosomes, which can be used for downstream analysis.
Protocol
1. Magnetic Bead-based Exosome Extraction Pipette a well-mixed solution of 5 µL of Streptavidin-coated magnetic microparticles into 495 µL of phosphate-buffered saline (PBS) buffer in a microcentrifuge tube to resuspend the beads. Wash and resuspend the beads with 500 µL of PBS three times using a magnetic rack. The rack is an array of magnets on the side of a housing unit that can hold the sample microcentrifuge tubes. For each wash, first, let the tubes sit on the…
Declarações
The authors have nothing to disclose.
Materials
| Biotinylated anti-human CD63 Antibody | Ancell, USA | 215-030 | |
| Dynabeads MyOne Streptavidin T1 | Invitrogen, USA | 65601 | |
| Neodynium Magnetics (1/10" dia. x 1/32" thick) | K&J Magnetics, USA | DH101 | |
| Ultrapure Distilled Water | Life Technologies, USA | 10977-023 | |
| Anti-Fluorescein-POD, Fab fragments | Roche, Germany | 11426346910 | |
| Phosphate Buffered Saline Solution | Life Technologies, USA | 10010023 | |
| Casein/PBS | Fisher Scientific, USA | 37532 |
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Magnetic Bead-based Exosome Extraction: A Technique to Isolate Specific Exosomes from Biofluids. J. Vis. Exp. (Pending Publication), e20496, doi: (2023).