Related Experiment Video
Updated: May 7, 2026
10:56
Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale
Published on: May 17, 2014
Polysome analysis of mammalian cells.
1Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Methods in Enzymology
|September 17, 2013
Summary
This study outlines a method to measure global translation in mammalian cells. It provides a protocol for analyzing polysomes to understand protein synthesis levels.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Translational control is crucial for cellular function and response to stimuli.
- Understanding global translation levels in mammalian cells is essential for various research areas.
- Existing methods for analyzing translation are often specific to non-mammalian systems.
Purpose of the Study:
- To establish a standardized protocol for assessing the global translational efficiency in mammalian cells.
- To provide a method for polysome profiling applicable to mammalian systems.
- To facilitate the study of mRNA and ribosome dynamics during protein synthesis.
Main Methods:
- The study details a protocol for polysome profiling in mammalian cells.
- This involves cell lysis, RNA extraction, and separation of polysomes via sucrose density gradient ultracentrifugation.
- Analysis of fractions to quantify mRNA and ribosome distribution.
Main Results:
- The protocol enables quantitative assessment of global translational activity.
- It allows for the characterization of polysome profiles specific to mammalian cells.
- The method can be adapted to study translational regulation under different conditions.
Conclusions:
- This protocol provides a valuable tool for researchers studying protein synthesis in mammalian cells.
- It offers a standardized approach to measure global translation, aiding comparative studies.
- The findings contribute to a deeper understanding of translational regulation in eukaryotes.
