Measuring Protein Synthesis in Cultured Cells and Mouse Tissues Using the Non-radioactive SUnSET Assay

Ravi, V and Jain, A and Mishra, S and Sundaresan, NR (2020) Measuring Protein Synthesis in Cultured Cells and Mouse Tissues Using the Non-radioactive SUnSET Assay. In: Current protocols in molecular biology, 133 (1).

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Abstract

Changes in protein synthesis occur under diverse physiological and pathological conditions. For example, translation can increase in response to growth signals or decrease in response to pathological states. Such changes have traditionally been measured by tracking the incorporation of radiolabeled amino acids. However, use of radioactivity is increasingly disfavored, and a simple and efficient puromycin-based, non-radioactive method called the SUnSET assay has gained popularity for measuring protein synthesis in diverse cell types and tissues. Here, we describe the principles, procedures, and troubleshooting steps for measuring protein synthesis using the SUnSET assay in cultured cells and mouse tissues. © 2020 Wiley Periodicals LLC Basic Protocol 1: Measuring protein synthesis in cultured cells by western blotting Support Protocol 1: Ponceau staining Support Protocol 2: Testing the specificity of the anti-puromycin antibody Basic Protocol 2: Measuring protein synthesis in cultured cells by immunofluorescence Basic Protocol 3: Measuring protein synthesis in mouse tissues by western blotting. © 2020 Wiley Periodicals LLC.

Item Type:	Journal Article
Publication:	Current protocols in molecular biology
Publisher:	NLM (Medline)
Additional Information:	Copyright to this article belongs to Blackwell Publishing Inc.
Department/Centre:	Division of Biological Sciences > Microbiology & Cell Biology Division of Interdisciplinary Sciences > Centre for Biosystems Science and Engineering
Date Deposited:	08 Jan 2021 06:44
Last Modified:	08 Jan 2021 06:44
URI:	http://eprints.iisc.ac.in/id/eprint/67168

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