2. Academic Validation
  3. Protocol for visualizing newly synthesized proteins in primary mouse hepatocytes

Protocol for visualizing newly synthesized proteins in primary mouse hepatocytes

  • STAR Protoc. 2021 Jun 17;2(3):100616. doi: 10.1016/j.xpro.2021.100616.
Yuqian Shen 1 2,  Wenhua Liu 1,  Jian Zuo 1,  Junhai Han 1 2,  Zi Chao Zhang 1 2
Affiliations

Affiliations

  • 1 School of Life Science and Technology, the Key Laboratory of Developmental Genes and Human Disease, Southeast University, Nanjing 210096, China.
  • 2 Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu 226001, China.
PMID: 34189479 DOI: 10.1016/j.xpro.2021.100616
Abstract

Selective identification of newly synthesized proteins is challenging because all proteins, both existing and nascent, have the same amino acid pool and are therefore chemically indistinguishable. L-homopropargylglycine is an amino acid analog of methionine containing an alkyne moiety that can undergo a classic click chemical reaction with Azide containing Alexa Fluor. Here, we present an integrated tool based on immunofluorescence staining to accurately trace and localize the newly synthesized protein in isolated primary mouse hepatocytes. For complete details on the use and execution of this protocol, please refer to Shen et al. (2021).

Keywords

Cell culture; Cell isolation; Cell-based Assays; Gene Expression; Microscopy; Molecular/Chemical Probes.

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