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Complementation of Splicing Activity by a Galectin-3 – U1 snRNP Complex on Beads
JoVE Journal
Biochimica
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JoVE Journal Biochimica
Complementation of Splicing Activity by a Galectin-3 – U1 snRNP Complex on Beads
DOI:

08:48 min

December 09, 2020

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Capitoli

  • 00:04Introduction
  • 00:53Preparation of NE Depleted of U1 snRNP (U1ΔNE)
  • 02:10Immunoprecipitation of 10S Fractions of Glycerol Gradients by Anti-Gal3
  • 03:52Assembly of Splicing Reaction and Analysis of Products
  • 06:09Results: RNA and Polypeptide Compositions of the Anti-Gal3 Precipitate of the 10S Region of the Glycerol Gradient and its Ability to Reconstitute Splicing in U1ΔNE
  • 08:08Conclusion

Summary

Traduzione automatica

This article describes the experimental procedures for (a) depletion of U1 snRNP from nuclear extracts, with concomitant loss of splicing activity; and (b) reconstitution of splicing activity in the U1-depleted extract by galectin-3 - U1 snRNP particles bound to beads covalently coupled with anti-galectin-3 antibodies.

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