Utilization of Grafix for the Detection of Transient Interactors of <em>Saccharomyces cerevisiae</em> Spliceosome Subcomplexes

Felipe A. Carvalho; Moises R. A. Barros; Thierry P. F. Girotto; M. Griselda Perona; Carla C. Oliveira

doi:10.3791/61994

Journal

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Biochimie

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

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Biochimie

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Journal JoVE Biochimie

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

•

November 09, 2020

DOI:

10.3791/61994-v

DOI:

10.3791/61994-v

•

05:44 min

November 09, 2020

•

8 Views

Felipe A. Carvalho¹, Moises R. A. Barros¹, Thierry P. F. Girotto¹, M. Griselda Perona¹, Carla C. Oliveira¹

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

Summary

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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Grafix Transient Interactors Saccharomyces Cerevisiae Spliceosome Subcomplexes Gradient Fixation Method Glycerol Gradient Centrifugation Cross-linker Fixation Reagents Yeast Total Extract Preparation Tap Tag Glycerol Gradient Glutaraldehyde Gradient Master Device

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

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