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Ribo Mega-SEC Provides a Rapid, Efficient and Reproducible Analysis of Mammalian Polysomes and Ribosomal Subunits

ABSTRACT: We describe Ribo Mega-SEC, a powerful approach for the separation and biochemical analysis of mammalian polysomes and ribosomal subunits using Size Exclusion Chromatography and uHPLC, which was achieved within 15 min from sample injection to fraction collection. Ribo Mega-SEC reproducibly shows translating ribosomes exist predominantly in polysome complexes in extracts isolated from human cell lines and mouse liver tissue, which alter in response to starvation. Ribo Mega-SEC provides a rapid, efficient, convenient and highly reproducible method for studying functional translation complexes and is easily combined with high-through put analysis such as proteomics and RNA-Seq, or with structural analysis using electron microscopy. We propose that Ribo Mega-SEC analysis is an accessible alternative to traditional polysome profiling using sucrose density gradients.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver

SUBMITTER: Mark Larance

LAB HEAD: Mark Larance

PROVIDER: PXD008913 | Pride | 2018-08-10

REPOSITORIES: Pride

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			Action	DRS
	20170728_QEPlus_ML_DH_EXP57_Polysomes_M1_FRAC01.raw	Raw
	20170728_QEPlus_ML_DH_EXP57_Polysomes_M1_FRAC02.raw	Raw
	20170728_QEPlus_ML_DH_EXP57_Polysomes_M1_FRAC03.raw	Raw
	20170728_QEPlus_ML_DH_EXP57_Polysomes_M1_FRAC04.raw	Raw
	20170728_QEPlus_ML_DH_EXP57_Polysomes_M1_FRAC05.raw	Raw

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Ribo Mega-SEC Provides a Rapid, Efficient and Reproducible Analysis of Mammalian Polysomes and Ribosomal Subunits

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