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Salavessa, Laura; Sauvonnet, Nathalie
Methods in molecular biology (Clifton, N.J.), 2021, Letnik: 2233Journal Article
Determination of protein stoichiometry in living cells is key to understanding basic biological processes. This is particularly important for receptor-mediated endocytosis, a highly regulated mechanism that requires the sequential assembly of numerous factors. Here, we describe a quantitative approach to analyze receptor clustering dynamics at the plasma membrane. Our workflow combines TIRF live imaging of a CRISPR-Cas9-edited cell line expressing a GFP-tagged receptor in a physiological relevant environment, a calibration technique for single-molecule analysis of GFP, and detection and tracking with an open-source software. This method allows to determine the number of receptor molecules at the plasma membrane in real time.
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| Leto | Faktor vpliva | Izdaja | Kategorija | Razvrstitev | ||||
|---|---|---|---|---|---|---|---|---|
| JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP | |
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in: SICRIS
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