☆ 4.5 Article

Filtration assay for arachidonic acid release

ANALYTICAL BIOCHEMISTRY (2010)

Journal

ANALYTICAL BIOCHEMISTRY

Volume 407, Issue 2, Pages 233-236

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE

DOI: 10.1016/j.ab.2010.07.014

Keywords

Arachidonic acid; Phospholipase A(2); Lipid messengers; G-protein-coupled receptor signaling

Categories

Biochemical Research Methods Biochemistry & Molecular Biology Chemistry, Analytical

Funding

Academy of Finland
Sigrid Juselius Foundation
Magnus Ehrnrooth Foundation
University of Helsinki

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Abstract

Arachidonic acid (AA) release is a central message in cell signaling. Fatty acid release is generally assessed by manual sampling of radioactivity release from cells prelabeled with a radiolabeled fatty acid. The assay is laborious, time-consuming, and susceptible to high noise. Here we present a fast and reproducible method for 96-well filter plates and cells in suspension, a method that is best suited for agonist concentration-response studies and, thus, for ligand screening. The method offers tremendous time and effort savings and enables execution of large experimental series previously unattainable for AA release studies. (C) 2010 Elsevier Inc. All rights reserved.

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