Measurement of Phospholipid Metabolism in Intact Neutrophils

  • Susan Sergeant
  • Linda C. McPhail
Part of the Methods in Molecular Biology book series (MIMB, volume 1124)


Phospholipid-metabolizing enzymes are important participants in neutrophil signal transduction pathways. The methods discussed herein describe assays for assessing the activities of phospholipase A2 (PLA2), phospholipase C (PLC), phospholipase D (PLD), and phosphoinositide 3-OH-kinase in intact neutrophils. PLA2 activity is measured as the release of radiolabeled 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.


Neutrophil Phospholipase Lipid kinase Phospholipid Signal transduction Lipid second messengers 



Development of the phospholipase D assay was partially supported by National Institutes of Health grant R01 AI-22564 to L.C.M.


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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Susan Sergeant
    • 1
  • Linda C. McPhail
    • 1
  1. 1.Department of BiochemistryWake Forest University School of MedicineWinston-SalemUSA

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