Journal
/
Biochemistry
/
Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
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

•

05:44 min

•

November 09, 2020

•

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

Chapters

  • 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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

Study on the Metabolism of Six Systemic Insecticides in a Newly Established Cell Suspension Culture Derived from Tea (Camellia Sinensis L.) Leaves

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Labelling and Visualization of Mitochondrial Genome Expression Products in Baker’s Yeast Saccharomyces cerevisiae

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Analysis of Neutral Lipid Synthesis in Saccharomyces cerevisiae by Metabolic Labeling and Thin Layer Chromatography

Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography

Assessment of Submitochondrial Protein Localization in Budding Yeast Saccharomyces cerevisiae

Purification of Endogenous Drosophila Transient Receptor Potential Channels

Read Article