Protocol for synthesis and use of a turn-on fluorescent probe for quantifying labile Zn2+ in the Golgi apparatus in live cells

Toshiyuki Kowada, Tomomi Watanabe, Rong Liu, Shin Mizukami

Research output: Contribution to journalArticlepeer-review

Abstract

Quantitative analysis using a turn-on fluorescent probe is inherently difficult due to the dependency of the fluorescence intensity on the probe concentration. To overcome this limitation, we developed an in situ quantification method using a turn-on fluorescent probe and a standard fluorophore, which are colocalized by protein tag technology. This protocol describes the synthesis of a Zn2+ probe, named ZnDA-1H, and the procedure to quantify the labile Zn2+ concentration in the Golgi of live HeLa cells by confocal fluorescence microscopy.

Original languageEnglish
Article number100395
JournalSTAR Protocols
Volume2
Issue number2
DOIs
Publication statusPublished - 2021 Jun 18

Keywords

  • Cell Biology
  • Cell-based Assays
  • Microscopy
  • Molecular/Chemical Probes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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