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Protocol for visualizing newly synthesized proteins in primary mouse hepatocytes.

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).

SUBMITTER: Shen Y

PROVIDER: S-EPMC8220402 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Protocol for visualizing newly synthesized proteins in primary mouse hepatocytes.

Shen Yuqian Y Liu Wenhua W Zuo Jian J Han Junhai J Zhang Zi Chao ZC

STAR protocols 20210617 3

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 ...[more]

PMID: 34189479

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