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

STAR protocols (2020-10-29)
Hiroyuki Kojima, Toshimasa Itoh
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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly-L-lysine hydrobromide, mol wt 30,000-70,000, lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture