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Fractionation of Subcellular Compartments from Human Brain Tissue

  • Toni M. MuellerEmail author
  • Pitna Kim
  • James H. Meador-Woodruff
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1941)

Abstract

Subcellular fractionation methods permit the isolation, purification, and/or enrichment of specific cellular compartments from complex tissue samples. Enrichment of multiple subcellular compartments from the same tissue sample permits comparisons of the spatial distribution of target proteins between specific intracellular compartments and, in some cases, can provide information about spatiotemporal processing of key cellular components. Here we describe a method to generate subcellular fractions enriched for heavy membranes and nuclei, rough and smooth endoplasmic reticulum membranes, light membranes and cytosol, synapses, and other intermediate cellular membranes from postmortem human brain tissue. These subcellular fractions can be used in a variety of downstream applications to assess the localization, relative abundance, and stoichiometry of glutamate receptor subunits along the forward trafficking pathway.

Key words

Density gradient separation Differential centrifugation Triton X-100 solubilization Forward trafficking pathway Subcellular localization 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Toni M. Mueller
    • 1
    Email author
  • Pitna Kim
    • 1
  • James H. Meador-Woodruff
    • 1
  1. 1.Department of Psychiatry and Behavioral NeurobiologyUniversity of Alabama at BirminghamBirminghamUSA

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