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Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate

DOI:
10.3791/59573-v

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11:13 min

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May 09, 2019

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, , , ,
1Center for Pathobiochemistry and Genetics, Institute of Medical Chemistry and Pathobiochemistry,Medical University Vienna, 2School of Medical Engineering and Applied Social Sciences,University of Applied Sciences Upper Austria

Chapters

  • 00:04Title
  • 00:35Reconstitution of HDL (High-Density Lipoprotein) Particles
  • 05:45Labeling of LDL (Low-Density Lipoprotein) Particles
  • 07:19Quality Control of Reconstituted/Labeled Lipoprotein Particles
  • 09:19Results
  • 10:34Conclusion

Summary

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Here, a quantitative real-time polymerase chain reaction-based protocol is presented for the determination of the native micro-RNA content (absolute/relative) of lipoprotein particles. In addition, a method for increasing the micro-RNA level, as well as a method for determining the cellular uptake rate of lipoprotein particles, is demonstrated.

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Keywords: Lipoprotein ParticlesMicroRNACellular UptakeDelipidationEnrichmentRelipidationBradford Protein AssayReconstitutionSpermineSynthetic MicroRNA

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