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Påvisning af RNA-bindende proteiner ved<em> In vitro</em> RNA Pull-down i fedtcellerne Kultur
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JoVE Journal Biology
Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

Påvisning af RNA-bindende proteiner ved<em> In vitro</em> RNA Pull-down i fedtcellerne Kultur

DOI:
10.3791/54207-v

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10:34 min

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July 22, 2016

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, , ,
1Cardiovascular and Metabolic Disorders Program,Duke-NUS Medical School, 2Institute of Molecular and Cell Biology, Singapore, 3Division of Bioengineering, School of Chemical & Biomedical Engineering,Nanyang Technological University

Chapters

  • 00:05Title
  • 00:35Preparation of Biotin-labeled RNA
  • 02:18Preparation of RNA-conjugated Beads
  • 04:16Preparation of Cellular Lysate from Primary Adipocytes
  • 07:00Binding and Elution of RBP
  • 08:45Results: RNA Pull-down Assay
  • 10:05Conclusion

Summary

Automatic Translation

Automatic Translation

En RNA pull-down protokol er optimeret her til påvisning af interaktioner mellem RNA-bindende proteiner (RBP'er) og ikke-kodende samt kodende RNA'er. En RNA-fragment fra androgen receptor (AR) blev anvendt som et eksempel for at vise hvordan man kan hente sin RBP fra lystate af primære brune fedtceller.

Tags

RNA-binding ProteinsRNA Pull-downAdipocyte CultureAntigen Receptor 3-Prime Untranslated Region (AR 3-Prime UTR)Biotinylated RNAIn Vitro TranscriptionStreptavidin-coated Magnetic BeadsRNA Structure BufferRNA-protein InteractionsRNA Regulatory MechanismsLong Non-coding RNA

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