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

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 ch...

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Bibliographic Details
Main Authors: Claire E. Dudley, Lotte van den Goor, Ann L. Miller
Format: Article
Language:English
Published: Elsevier 2022-09-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166722005020
Description
Summary: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.
ISSN:2666-1667