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  • Immobilization of bacterial luciferase into poly(N-isopropylacrylamide) film for electrochemical control of a bioluminescence reaction.

Immobilization of bacterial luciferase into poly(N-isopropylacrylamide) film for electrochemical control of a bioluminescence reaction.

Analytical sciences : the international journal of the Japan Society for Analytical Chemistry (2012-10-13)
Yoji Kawanami, Shinya Yamasaki, Shuto Yamada, Kô Takehara
ABSTRACT

A poly(N-isopropylacrylamide) film was modified on an indium-tin oxide electrode in order to immobilize bacterial luciferase (BL) on the electrode surface. By using the modified electrode, flavin mononucleotide (FMN) was electrochemically reduced to FMNH(2), which is one of the substrates of the BL luminescence reaction, to control the bioluminescence reaction by BL. The BL reaction in the modified film could be promoted and controlled by the electrochemical generation of FMNH(2). This BL luminescence system was evaluated as a model system for the inhibitory assay of hydrophobic molecules on protein functions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly(N-isopropylacrylamide), carboxylic acid terminated, average Mn 10,000
Sigma-Aldrich
Poly(N-isopropylacrylamide), Mn ~40,000
Sigma-Aldrich
Lauryl alcohol, ≥98%, FG
Supelco
1-Dodecanol, Selectophore, ≥98.0%
Sigma-Aldrich
1-Dodecanol, reagent grade, 98%
Sigma-Aldrich
1-Dodecanol, ACS reagent, ≥98.0%
Sigma-Aldrich
Poly(N-isopropyl acrylamide), NHS ester end functionalized, average Mn 5,000