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Measurement of Phospholipid Metabolism in Intact Neutrophils

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Neutrophil Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1124))

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Abstract

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.

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Acknowledgements

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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Authors and Affiliations

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

    Susan Sergeant & Linda C. McPhail

Authors
  1. Susan Sergeant
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  2. Linda C. McPhail
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Editor information

Editors and Affiliations

  1. Montana State University Dept. Veterinary Molecular Biology, Bozeman, Montana, USA

    Mark T. Quinn

  2. Lab. Human Bacterial Pathogenesis, National Institutes of Health Nat. Inst. Allergy & Infectious Diseases, Hamilton, Montana, USA

    Frank R. DeLeo

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Sergeant, S., McPhail, L.C. (2014). Measurement of Phospholipid Metabolism in Intact Neutrophils. In: Quinn, M., DeLeo, F. (eds) Neutrophil Methods and Protocols. Methods in Molecular Biology, vol 1124. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-845-4_7

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  • DOI: https://doi.org/10.1007/978-1-62703-845-4_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-844-7

  • Online ISBN: 978-1-62703-845-4

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