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Analysis of local protein accumulation kinetics by live-cell imaging in yeast systems.

STAR protocols (2021-08-31)
Hiroki Okada, Brittany MacTaggart, Erfei Bi
ABSTRACT

Microscopy-based analysis of protein accumulation at a given subcellular location in real time provides invaluable insights into the function of a protein in a specific process. Here, we describe a detailed protocol for determining protein accumulation kinetics at the division site in the budding yeast Saccharomyces cerevisiae and fission yeast Schizosaccharomyces pombe. This protocol can be adapted for the analysis of any protein involved in any process as long as the protein is localized to a discrete region of the cell. For complete details on the use and execution of this protocol, please refer to Okada et al. (2021) and Okada et al. (2019).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Lectin from Glycine max (soybean), lyophilized powder, salt, essentially free
Sigma-Aldrich
Uracil, ≥99.0%
Sigma-Aldrich
myo-Inositol, ≥99%
Sigma-Aldrich
Concanavalin A from Canavalia ensiformis (Jack bean), Type VI, lyophilized powder