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Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells
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Cancer Research
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JoVE Journal Cancer Research
Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells

Desthiobiotin-Streptavidin-Affinity Mediated Purification of RNA-Interacting Proteins in Mesothelioma Cells

DOI:
10.3791/57516-v

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11:11 min

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April 25, 2018

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,
1Laboratory of Molecular Oncology, Division of Thoracic Surgery,University Hospital Zurich

Chapters

  • 00:04Title
  • 00:49Cytosolic and Nuclear Protein Fraction Preparation
  • 03:52Labeling RNA with Desthiobiotin
  • 06:04RNA-Protein Pulldown
  • 09:24Results: High Specificity Binding Via Protocol
  • 10:33Conclusion

Summary

Automatic Translation

Automatic Translation

Desthiobiotin labeling of a synthetic 25-nucleotide RNA oligo, which contains an adenine-rich element (ARE) motif, allows specific binding of cytosolic ARE-binding protein.

Tags

Keywords: DesthiobiotinStreptavidinAffinity PurificationRNA-interacting ProteinsMesothelioma CellsCytosolic ProteinRNA BindingReporter AssayACC-MESO-4 CellsTrypsin-EDTARPMI-1640 MediumPBSCytoplasmic ExtractionProteinase InhibitorNuclear Extraction

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