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Analysis of Fatty Acid and Cholesterol Content from Detergent-Resistant and Detergent-Free Membrane Microdomains

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Book cover Lipidomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1609))

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Abstract

The compartmentalization of cellular membranes into discrete membrane microdomains (known as lipid rafts) challenged the original definition of membranes as containing randomly distributed lipid and protein components. The lipid microdomain hypothesis has generated significant controversy and rigorous inquiry based on the attractive idea that such domains concentrate machinery to mediate cellular events such as signaling and endocytosis. As such, numerous studies have used biochemical, cell biological, and biophysical methodologies to define the composition of such domains in a variety of experimental contexts. In this chapter, we describe methodologies to isolate membranes from cell or tissue sources with biophysical/biochemical properties of membrane microdomains that are amenable to subsequent classical or mass spectrometry-based lipid analytical approaches.

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Acknowledgments

This work was supported by NIH Grant R01EY019494, NIH Core Grant P30EY021725, and by an unrestricted grant from Research to Prevent Blindness, Inc. to the Department of Ophthalmology, University of Oklahoma Health Science Center.

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Authors and Affiliations

  1. Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA

    Mark E. McClellan & Michael H. Elliott

  2. Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA

    Michael H. Elliott

  3. Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA

    Michael H. Elliott

Authors
  1. Mark E. McClellan
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  2. Michael H. Elliott
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Corresponding author

Correspondence to Michael H. Elliott .

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Editors and Affiliations

  1. University of Miami, Miami, Florida, USA

    Sanjoy K. Bhattacharya

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McClellan, M.E., Elliott, M.H. (2017). Analysis of Fatty Acid and Cholesterol Content from Detergent-Resistant and Detergent-Free Membrane Microdomains. In: Bhattacharya, S. (eds) Lipidomics. Methods in Molecular Biology, vol 1609. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6996-8_16

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  • DOI: https://doi.org/10.1007/978-1-4939-6996-8_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6995-1

  • Online ISBN: 978-1-4939-6996-8

  • eBook Packages: Springer Protocols

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