SNAP- and Halo-tagging and dye introduction protocol for live microscopy in Xenopus embryos

Autor: Claire E. Dudley, Lotte van den Goor, Ann L. Miller
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: STAR Protocols, Vol 3, Iss 3, Pp 101622- (2022)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2022.101622
Popis: Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Databáze: Directory of Open Access Journals