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Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture
JoVE Journal
Biologia
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JoVE Journal Biologia
Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture
DOI:

10:34 min

July 22, 2016

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Capítulos

  • 00:05Título
  • 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

Tadução automática

An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.

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