Preparation and Use of Turn-on Fluorescent Probe for Detection and Live Cell Imaging of Vitamin D Receptor as a Target Protein

STAR Protoc. 2020 Jun 3;1(1):100036. doi: 10.1016/j.xpro.2020.100036. eCollection 2020 Jun 19.

Abstract

Turn-on fluorescent probe mediated by conjugate addition and cyclization (TCC probe) is a small molecule that reacts with a protein of interest in cells. TCC probe is applicable to various types of proteins by exchanging the ligand unit for target proteins. TCC probes are a potent tool for molecular imaging and chemical proteomics. This protocol describes the synthesis of a TCC probe via unstable intermediate and how to use this probe to visualize vitamin D receptor as a target protein. For complete details on the use and execution of this protocol, please refer to Kojima et al. (2020).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Dyes* / chemical synthesis
  • Fluorescent Dyes* / chemistry
  • Fluorescent Dyes* / metabolism
  • HeLa Cells
  • Humans
  • Molecular Imaging / methods*
  • Optical Imaging / methods*
  • Receptors, Calcitriol* / analysis
  • Receptors, Calcitriol* / chemistry
  • Receptors, Calcitriol* / metabolism

Substances

  • Fluorescent Dyes
  • Receptors, Calcitriol
  • VDR protein, human