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  • Extraction and quantification of adenosine triphosphate in mammalian tissues and cells.

Extraction and quantification of adenosine triphosphate in mammalian tissues and cells.

Methods in molecular biology (Clifton, N.J.) (2013-10-30)
Junji Chida, Hiroshi Kido
ABSTRACT

Adenosine 5'-triphosphate (ATP) is the "energy currency" of organisms and plays central roles in bioenergetics, whereby its level is used to evaluate cell viability, proliferation, death, and energy transmission. In this chapter, we describe an improved and efficient method for extraction of ATP from tissues and cells using phenol-based reagents. The chaotropic extraction reagents reported so far co-precipitate ATP with insoluble proteins during extraction and with salts during neutralization. In comparison, the phenol-based reagents extract ATP well without the risks of co-precipitation. The extracted ATP can be quantified by the luciferase assay or high-performance liquid chromatography.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Luciferase from Photinus pyralis (firefly), recombinant, expressed in E. coli, buffered aqueous solution, ≥10×1010 units/mg protein