Chunk #6 — Methods — Membrane Preparation

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Suppression of amygdalar endocannabinoid signaling by stress contributes to activation of the hypothalamic-pituitary-adrenal axis.

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Brain sections were homogenized in 10 volumes of TME buffer (50 mM Tris HCl, pH 7.4; 1 mM EDTA and 3 mM MgCl2). Homogenates were centrifuged at 18,000 × g for 20 min and the resulting pellet, which constituted the membrane fraction, was resuspended in 10 volumes of TME buffer. Protein concentrations were determined by the Bradford method (Bio-Rad, Hercules, CA, USA).