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Enhanced firefly bioluminescent assay of adenosine 5′‐triphosphate using liposomes containing cationic cholesterols
Author(s) -
Nakata Naohiro,
Kamidate Tamio
Publication year - 2001
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/bio.648
Subject(s) - bioluminescence , firefly protocol , liposome , chemistry , luciferase , adenosine triphosphate , luminescent measurements , cationic liposome , cationic polymerization , luciferases , biochemistry , biology , transfection , physics , luminescence , zoology , organic chemistry , gene , optoelectronics
Cationic liposomes composed of phosphatidylcholine and cationic cholesterols were prepared by extrusion technique. Dimethylaminoethyl‐carbamoyl cholesterol (DMAE‐chol) and diethylaminoethyl‐carbamoyl cholesterol (DEAE‐chol) were synthesized as a cationic cholesterol. Cationic liposomes containing DMAE‐chol and DEAE‐chol enhanced the intensity of maximum light emission from the firefly bioluminescent (BL) reaction. The sensitivity for ATP in the presence of cationic liposomes containing DMAE‐chol and DEAE‐chol was improved by a factor of 10 times compared to that in water alone. The detection limit for ATP upon using cationic liposomes was 1.0 pmol/L in an aqueous standard solution. The BL enhancement in the presence of cationic liposomes could be explained in terms of BL emitters and electrostatic interaction between the liposomes surface and BL reactants. Copyright © 2001 John Wiley & Sons, Ltd.