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Co‐determination of ATP and proteins in Triton X 100 non‐ionic detergent‐opened monolayer cultured cells
Author(s) -
Kőszegi Tamás,
Petrik József,
VladimirKnežević Sanda,
Nagy Sándor
Publication year - 2007
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.979
Subject(s) - monolayer , chemistry , chromatography , biochemistry , extraction (chemistry) , intracellular , cell
Human monolayer cells (HEp‐2 and Hep G2) were cultured in 96‐well plates. A modified Triton X 100 nonionic detergent extraction method was used for releasing intracellular ATP and protein in one step. The detergent technique was compared to perchloric acid (PCA) extraction. ATP was determined by the firefly bioluminescence method and ATP values were referred to cell protein (ATP:protein ratio). There was no significant difference in ATP data between detergent and PCA treatments. The ATP:protein ratio seems to be a sensitive tool for characterizing the metabolic activity of monolayer tissue culture cells. The protein‐mobilizing capability of Triton X 100 depends on the type of cell culture used. Our modified extraction gives reliable ATP:protein values with one simple extraction step. Copyright © 2007 John Wiley & Sons, Ltd.