Enhanced fluorescent imaging of proteins in live yeast cells using fluorescently labeled scFv

Ioannis Tsirkas; Tomer Zur; Daniel Dovrat; Zohar Paleiov; Lior Ravkaie; Amir Aharoni

doi:10.1016/j.xpro.2023.102299

Enhanced fluorescent imaging of proteins in live yeast cells using fluorescently labeled scFv

Ioannis Tsirkas, Tomer Zur, Daniel Dovrat, Zohar Paleiov, Lior Ravkaie, Amir Aharoni

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

Fluorescent labeling of proteins is a widespread approach for the microscopic examination of protein function, expression, and localization in the cell. Here, we present a protocol for the labeling of hemagglutinin (HA)-tagged protein of interest (POI) with the single-chain antibody (scFv) 2E2 fused to different fluorescent proteins (FPs) in Saccharomyces cerevisiae. We describe steps for expressing 2E2-FP, and HA tagging and labeling of POI. We detail in vivo fluorescent imaging of proteins at different cellular compartments and with diverse expression levels. For complete details on the use and execution of this protocol, please refer to Tsirkas et al. (2022).¹

Original language	English
Article number	102299
Journal	STAR Protocols
Volume	4
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2023.102299
State	Published - 16 Jun 2023

Keywords

Cell Biology
Microscopy
Molecular/Chemical Probes
Single Cell

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2023.102299

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abstract = "Fluorescent labeling of proteins is a widespread approach for the microscopic examination of protein function, expression, and localization in the cell. Here, we present a protocol for the labeling of hemagglutinin (HA)-tagged protein of interest (POI) with the single-chain antibody (scFv) 2E2 fused to different fluorescent proteins (FPs) in Saccharomyces cerevisiae. We describe steps for expressing 2E2-FP, and HA tagging and labeling of POI. We detail in vivo fluorescent imaging of proteins at different cellular compartments and with diverse expression levels. For complete details on the use and execution of this protocol, please refer to Tsirkas et al. (2022).1",

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AU - Tsirkas, Ioannis

AU - Zur, Tomer

AU - Dovrat, Daniel

AU - Paleiov, Zohar

AU - Ravkaie, Lior

AU - Aharoni, Amir

PY - 2023/6/16

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AB - Fluorescent labeling of proteins is a widespread approach for the microscopic examination of protein function, expression, and localization in the cell. Here, we present a protocol for the labeling of hemagglutinin (HA)-tagged protein of interest (POI) with the single-chain antibody (scFv) 2E2 fused to different fluorescent proteins (FPs) in Saccharomyces cerevisiae. We describe steps for expressing 2E2-FP, and HA tagging and labeling of POI. We detail in vivo fluorescent imaging of proteins at different cellular compartments and with diverse expression levels. For complete details on the use and execution of this protocol, please refer to Tsirkas et al. (2022).1

KW - Cell Biology

KW - Microscopy

KW - Molecular/Chemical Probes

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Enhanced fluorescent imaging of proteins in live yeast cells using fluorescently labeled scFv

Abstract

Keywords

ASJC Scopus subject areas

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