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Phosphoinositides pp 1–17Cite as

Simultaneous Detection of Phosphoinositide Lipids by Radioactive Metabolic Labeling

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2251))

Abstract

Phosphoinositide (PPI) lipids are a crucial class of low-abundance signaling molecules that regulate many processes within cells. Methods that enable simultaneous detection of all PPI lipid species provide a wholistic snapshot of the PPI profile of cells, which is critical for probing PPI biology. Here we describe a method for the simultaneous measurement of cellular PPI levels by metabolically labeling yeast or mammalian cells with myo-3H-inositol, extracting radiolabeled glycerophosphoinositides, and separating lipid species on an anion exchange column via HPLC.

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Acknowledgments

This work was supported by NIH grants R01-NS099340-03 and R01-NS064015-09 to LSW, and LSI Cubed to NS and SSPG. NS was supported in part by NIH T-32-GM007315.

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Author notes

  1. Noah Steinfeld and Sai Srinivas Panapakkam Giridharan contributed equally to this work.

Authors and Affiliations

  1. Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA

    Noah Steinfeld, Sai Srinivas Panapakkam Giridharan, Emily J. Kauffman & Lois S. Weisman

  2. Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, MI, USA

    Noah Steinfeld

  3. Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA

    Lois S. Weisman

Authors
  1. Noah Steinfeld
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  2. Sai Srinivas Panapakkam Giridharan
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  3. Emily J. Kauffman
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  4. Lois S. Weisman
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Corresponding author

Correspondence to Lois S. Weisman .

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Editors and Affiliations

  1. Department of Chemistry and Biology and the Graduate Program in Molecular Science, Ryerson University, Toronto, ON, Canada

    Roberto J. Botelho

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Steinfeld, N., Giridharan, S.S.P., Kauffman, E.J., Weisman, L.S. (2021). Simultaneous Detection of Phosphoinositide Lipids by Radioactive Metabolic Labeling. In: Botelho, R.J. (eds) Phosphoinositides. Methods in Molecular Biology, vol 2251. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1142-5_1

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  • DOI: https://doi.org/10.1007/978-1-0716-1142-5_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1141-8

  • Online ISBN: 978-1-0716-1142-5

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