Co-determination of ATP and proteins in Triton X 100 non-ionic detergent-opened monolayer cultured cells.
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.
Koszegi
Tamás
T
Institute of Laboratory Medicine, University of Pécs, Hungary. koszegit@freemail.hu
Petrik
József
J
Vladimir-Knezević
Sanda
S
Nagy
Sándor
S
D000255:Adenosine Triphosphate / Q000032:analysis* / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003902:Detergents / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008163:Luminescent Measurements / Q000379:methods; D017830:Octoxynol / Q000737:chemistry* / Q000494:pharmacology; D010472:Perchlorates / Q000737:chemistry / Q000494:pharmacology; D011506:Proteins / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism; D012680:Sensitivity and Specificity
D003902:Detergents; D010472:Perchlorates; D011506:Proteins; D000255:Adenosine Triphosphate; D017830:Octoxynol