Tissue processing prior to protein analysis and amyloid-beta quantitation

Methods Mol Biol. 2005:299:267-78. doi: 10.1385/1-59259-874-9:267.

Abstract

Amyloid-containing tissue, whether from human patients or an animal model of a disease, is typically characterized by various biochemical and immunohistochemical techniques, many of which are described in detail in this volume. In this chapter, we describe a straightforward technique for the homogenization of tissue prior to these analyses. The technique is particularly well-suited for performing a large number of different biochemical analyses on a single mouse brain hemisphere. Starting with this homogenate, multiple characterizations can be done, including Western blot analysis and isolation of membrane-associated proteins, both of which are described here. Additional analyses can readily be performed on the tissue homogenate, including the ELISA quantitation of Abeta in the brain of a transgenic mouse model of beta-amyloid deposition. The ELISA technique is described in detail in the following chapter.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amyloid beta-Peptides / analysis*
  • Amyloid beta-Peptides / isolation & purification*
  • Amyloid beta-Peptides / metabolism
  • Amyloid beta-Protein Precursor / analysis
  • Amyloid beta-Protein Precursor / isolation & purification
  • Amyloid beta-Protein Precursor / metabolism
  • Animals
  • Blotting, Western / methods
  • Brain / metabolism
  • Humans
  • Mice
  • Tissue Fixation / methods

Substances

  • Amyloid beta-Peptides
  • Amyloid beta-Protein Precursor