Click-iT (R) Plus OPP Alexa Fluor (R) Protein Synthesis Assay in Embryonic Cells

Li, Y; Ji, X; Chang, L; Tang, J; Hua, M-M; Liu, J; O'Neill, C; Huang, X; Jin, X

Click-iT (R) Plus OPP Alexa Fluor (R) Protein Synthesis Assay in Embryonic Cells

Li, Y Ji, X Chang, L Tang, J Hua, M-M Liu, J O'Neill, C Huang, X Jin, X

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Publisher:: BIO-PROTOCOL
Publication Type:: Journal Article
Citation:: BIO-PROTOCOL, 2022, 12, (11), pp. e4441
Issue Date:: 2022-06-01

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Field	Value	Language
dc.contributor.author	Li, Y
dc.contributor.author	Ji, X
dc.contributor.author	Chang, L
dc.contributor.author	Tang, J
dc.contributor.author	Hua, M-M
dc.contributor.author	Liu, J
dc.contributor.author	O'Neill, C
dc.contributor.author	Huang, X
dc.contributor.author	Jin, X
dc.date.accessioned	2023-06-17T00:42:37Z
dc.date.available	2022-05-07
dc.date.available	2023-06-17T00:42:37Z
dc.date.issued	2022-06-01
dc.identifier.citation	BIO-PROTOCOL, 2022, 12, (11), pp. e4441
dc.identifier.issn	2331-8325
dc.identifier.issn	2331-8325
dc.identifier.uri	http://hdl.handle.net/10453/170775
dc.description.abstract	This protocol describes a method to assess relative changes in the level of global protein synthesis in the preimplantation embryo using the Click-iT (R) Plus OPP Protein Synthesis Assays. In this assay, O-propargylpuromycin (OPP), an analog of puromycin, is efficiently incorporated into the nascent polypeptide of newly translated proteins in embryonic cells. OPP is fluorescently labeled with a photostable Alexa FluorTM dye and detected with fluorescence microscopy. The intensity of the fluorescence is quantitatively analyzed. This is a fast, sensitive, and non-radioactive method for the detection of protein synthesis in early embryo development. It provides a tool for analyzing the temporal regulation of protein synthesis, as well as the effects of changes in the embryonic microenvironment, and pharmacological and genetic modulations of embryo development.
dc.language	eng
dc.publisher	BIO-PROTOCOL
dc.relation.ispartof	BIO-PROTOCOL
dc.relation.isbasedon	10.21769/BioProtoc.4441
dc.rights	info:eu-repo/semantics/closedAccess
dc.title	Click-iT (R) Plus OPP Alexa Fluor (R) Protein Synthesis Assay in Embryonic Cells
dc.type	Journal Article
utslib.citation.volume	12
utslib.location.activity	United States
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
utslib.copyright.status	closed_access	*
dc.date.updated	2023-06-17T00:42:36Z
pubs.issue	11
pubs.publication-status	Published online
pubs.volume	12
utslib.citation.issue	11

Abstract:

This protocol describes a method to assess relative changes in the level of global protein synthesis in the preimplantation embryo using the Click-iT (R) Plus OPP Protein Synthesis Assays. In this assay, O-propargylpuromycin (OPP), an analog of puromycin, is efficiently incorporated into the nascent polypeptide of newly translated proteins in embryonic cells. OPP is fluorescently labeled with a photostable Alexa FluorTM dye and detected with fluorescence microscopy. The intensity of the fluorescence is quantitatively analyzed. This is a fast, sensitive, and non-radioactive method for the detection of protein synthesis in early embryo development. It provides a tool for analyzing the temporal regulation of protein synthesis, as well as the effects of changes in the embryonic microenvironment, and pharmacological and genetic modulations of embryo development.

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http://hdl.handle.net/10453/170775