Size-Exclusion Chromatography Separation Reveals That Vesicular and Non-Vesicular Small RNA Profiles Differ in Cell Free Urine
Abstract
:1. Introduction
2. Results
2.1. Comparison of Particle and Protein Yield between Methods
2.2. Transmission Electron Microscopy Reveals Heterogeneous Vesicle Populations
2.3. Enriched Extracellular Vesicles Contain Mainly 40–80 nt Small RNA
3. Discussion
4. Materials and Methods
4.1. Urine Collection and Pre-Processing
4.2. Ultracentrifugation
4.3. Precipitation
4.4. Ultrafiltration
4.5. Size-Exclusion Chromatography
4.6. Nanoparticle Tracking Analysis
4.7. Protein Concentration
4.8. Transmission Electron Microscopy
4.9. RNA Isolation and Analysis
4.10. Western Blotting
4.11. Statistical Analyses
4.12. EV-TRACK
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Karttunen, J.; Stewart, S.E.; Kalmar, L.; Grant, A.J.; Karet Frankl, F.E.; Williams, T.L. Size-Exclusion Chromatography Separation Reveals That Vesicular and Non-Vesicular Small RNA Profiles Differ in Cell Free Urine. Int. J. Mol. Sci. 2021, 22, 4881. https://doi.org/10.3390/ijms22094881
Karttunen J, Stewart SE, Kalmar L, Grant AJ, Karet Frankl FE, Williams TL. Size-Exclusion Chromatography Separation Reveals That Vesicular and Non-Vesicular Small RNA Profiles Differ in Cell Free Urine. International Journal of Molecular Sciences. 2021; 22(9):4881. https://doi.org/10.3390/ijms22094881
Chicago/Turabian StyleKarttunen, Jenni, Sarah E. Stewart, Lajos Kalmar, Andrew J. Grant, Fiona E. Karet Frankl, and Tim L. Williams. 2021. "Size-Exclusion Chromatography Separation Reveals That Vesicular and Non-Vesicular Small RNA Profiles Differ in Cell Free Urine" International Journal of Molecular Sciences 22, no. 9: 4881. https://doi.org/10.3390/ijms22094881