Preparation of Membrane Extracts from Plant Tissues.

The reproducible isolation of subcellular membrane systems is crucial to investigate organelle-specific metabolic processes or protein targeting in plant cells. This chapter describes a modular protocol for the sequential enrichment of multiple membrane-bound compartments applicable to different eudicots, such as Medicago × varia and Solanum lycopersicum. Exemplified by Arabidopsis thaliana, sequential organelle-specific centrifugation and density gradient separation are utilized to obtain each compartment from a single homogenate of whole leaf tissue. This enables comparative biochemical or proteomic analyses across compartments.