Quantification of Choline Release

1–50-μl samples were incubated with 1 mM LPC (from egg) in the presence of 100 mM Tris-HCl, pH 9.0, 500 mM NaCl, 5 mM MgCl₂, and 0.05% Triton X-100 for 1 h at 37°C. The liberated choline was detected by an enzymatic photometric method using choline oxidase (Asahi Chemical), horseradish peroxidase (Toyobo), and TOOS reagent (N-ethyl-N-(2-hydoroxy-3-sulfoproryl)-3-methylaniline; Dojindo Molecular Technologies, Inc.) as a hydrogen donor (Imamura and Horiuti, 1978 (link); Tamaoku et al., 1982 ).

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Umezu-Goto M., Kishi Y., Taira A., Hama K., Dohmae N., Takio K., Yamori T., Mills G.B., Inoue K., Aoki J, & Arai H. (2002). Autotaxin has lysophospholipase D activity leading to tumor cell growth and motility by lysophosphatidic acid production. The Journal of Cell Biology, 158(2), 227-233.

Publication 2002

Choline Choline oxidase Donor Enzymatic Horseradish peroxidase Hydrogen Methylaniline Mgcl2 Nacl Photometric method Toos Tris Triton x 100

Corresponding Organization :

Other organizations : The University of Tokyo, RIKEN, Japanese Foundation For Cancer Research, Center for Systems Biology, The University of Texas MD Anderson Cancer Center, Teikyo University

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Variable analysis

independent variables

LPC (from egg) concentration: 1 mM

dependent variables

Liberated choline concentration (detected by enzymatic photometric method)

control variables

Tris-HCl buffer concentration: 100 mM
NaCl concentration: 500 mM
MgCl2 concentration: 5 mM
Triton X-100 concentration: 0.05%
Incubation time: 1 h
Incubation temperature: 37°C

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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