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

3,316 Views

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05:44 min

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November 09, 2020

DOI:

10.3791/61994-v

DOI:
10.3791/61994-v

•

05:44 min
November 09, 2020

•

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

概要

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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.

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

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Cite this Article

Carvalho, F. A., Barros, M. R. A., Girotto, T. P. F., Perona, M. G., Oliveira, C. C. Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes. J. Vis. Exp. (165), e61994, doi:10.3791/61994 (2020).

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