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Methods Mol Biol. 2007;412:69-83. doi: 10.1007/978-1-59745-467-4_6.

Measurement of phospholipid metabolism in intact neutrophils.

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1
Department of Biochemistry, Wake Forest University, School of Medicine, Winston-Salem, NC, USA.

Abstract

Phospholipid metabolizing enzymes are important participants in neutrophil signal transduction pathways. The methods discussed herein describe assays for assessing the activities of phospholipase (PL)A2, PLC, PLD, and phosphoinositide 3-OH-kinase (PI3-K) in intact neutrophils. PLA2 activity is measured as the release of radiolabed arachidonic acid. PLC activity is measured as the accumulation of inositol 1,4,5-trisphosphate (IP3), a water-soluble product, using a commercially available radioreceptor assay kit. PLD activity is measured as the appearance of its radiolabeled products, phosphatidic acid and phosphatidylethanol. PI3-K activity is measured as the appearance of its radiolabeled product, phosphatidylinositol-3,4,5-trisphosphate.

PMID:
18453106
DOI:
10.1007/978-1-59745-467-4_6
[Indexed for MEDLINE]
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