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Amperometric Detection of Apoptosis by using p ‐Methoxyaniline‐Conjugated Substrate for Caspase‐3
Author(s) -
Sixiang Sun,
Inoue Kumi Y.,
Shiomoto Shusaku,
Takano Shinichiro,
Ino Kosuke,
Shiku Hitoshi,
Matsue Tomokazu
Publication year - 2017
Publication title -
chemelectrochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.182
H-Index - 59
ISSN - 2196-0216
DOI - 10.1002/celc.201600700
Subject(s) - amperometry , apoptosis , substrate (aquarium) , chronoamperometry , cyclic voltammetry , phosphate buffered saline , chemistry , cell culture , chromatography , lysis , detection limit , electrochemistry , biophysics , biochemistry , microbiology and biotechnology , biology , electrode , ecology , genetics
This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp‐Glu‐Val‐Asp‐ p ‐methoxyaniline (DEVD‐ p MA). This substrate showed electrochemical inertia from −0.2 to 0.7 V versus Ag/AgCl in phosphate‐buffered saline (PBS), but was cleaved by caspase‐3, releasing p MA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p ‐aminophenol ( p AP), p MA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase‐3 in the range of 0.31–2.5×10 −2 units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.