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SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos
Traditional fluorescent proteins exhibit limitations in brightness and photostability that hinder optimal characterization of the dynamic cellular behavior of proteins of interest. SNAP- and Halo-tagging are alternatives to traditional fluorescent protein tagging utilizing bright, stable chemical dy...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9405085/ https://www.ncbi.nlm.nih.gov/pubmed/36035797 http://dx.doi.org/10.1016/j.xpro.2022.101622 |
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author | Dudley, Claire E. van den Goor, Lotte Miller, Ann L. |
author_facet | Dudley, Claire E. van den Goor, Lotte Miller, Ann L. |
author_sort | Dudley, Claire E. |
collection | PubMed |
description | Traditional fluorescent proteins exhibit limitations in brightness and photostability that hinder optimal characterization of the dynamic cellular behavior of proteins of interest. SNAP- and Halo-tagging are alternatives to traditional fluorescent protein tagging utilizing bright, stable chemical dyes, which may improve signal-to-noise ratio. However, there has been limited use of this approach in vivo in developing organisms. Here, we present a protocol for implementing SNAP- and Halo-tagging in gastrula-stage Xenopus laevis embryos for live confocal microscopy. For complete details on the use and execution of this protocol, please refer to Varadarajan et al. (2022). |
format | Online Article Text |
id | pubmed-9405085 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-94050852022-08-26 SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos Dudley, Claire E. van den Goor, Lotte Miller, Ann L. STAR Protoc Protocol Traditional fluorescent proteins exhibit limitations in brightness and photostability that hinder optimal characterization of the dynamic cellular behavior of proteins of interest. SNAP- and Halo-tagging are alternatives to traditional fluorescent protein tagging utilizing bright, stable chemical dyes, which may improve signal-to-noise ratio. However, there has been limited use of this approach in vivo in developing organisms. Here, we present a protocol for implementing SNAP- and Halo-tagging in gastrula-stage Xenopus laevis embryos for live confocal microscopy. For complete details on the use and execution of this protocol, please refer to Varadarajan et al. (2022). Elsevier 2022-08-18 /pmc/articles/PMC9405085/ /pubmed/36035797 http://dx.doi.org/10.1016/j.xpro.2022.101622 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Dudley, Claire E. van den Goor, Lotte Miller, Ann L. SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos |
title | SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos |
title_full | SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos |
title_fullStr | SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos |
title_full_unstemmed | SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos |
title_short | SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos |
title_sort | snap- and halo-tagging and dye introduction protocol for live microscopy in xenopus embryos |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9405085/ https://www.ncbi.nlm.nih.gov/pubmed/36035797 http://dx.doi.org/10.1016/j.xpro.2022.101622 |
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