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Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes
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Bioquímica
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JoVE Journal Bioquímica
Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes

Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes

DOI:
10.3791/61994-v

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05:44 min

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November 09, 2020

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, , , ,
1Department of Biochemistry, Institute of Chemistry,University of Sao Paulo

Capítulos

  • 00:04Introduction
  • 00:45Yeast Total Extract Preparation
  • 02:07Glycerol Gradient Preparation
  • 02:58Extract Separation and Collection
  • 03:44Results: Representative Transient Saccharomyces Cerevisiae Spliceosome Subcomplex Interactor Detection
  • 05:15Conclusion

Summary

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Here, we describe the utilization of Grafix (Gradient Fixation), a glycerol gradient centrifugation in the presence of a crosslinker, to identify interactions between splicing factors that bind transiently to the spliceosome complex.

Tags

GrafixTransient InteractorsSaccharomyces CerevisiaeSpliceosome SubcomplexesGradient Fixation MethodGlycerol Gradient CentrifugationCross-linkerFixation ReagentsYeast Total Extract PreparationTap TagGlycerol GradientGlutaraldehydeGradient Master Device

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