A protocol for quantitative analysis of murine and human amyloid-β1-40 and 1-42

J Neurosci Methods. 2017 Nov 1:291:28-35. doi: 10.1016/j.jneumeth.2017.07.022. Epub 2017 Jul 30.

Abstract

Background: Amyloid-β (Aβ), a hallmark of Alzheimer's disease (AD), has long been a focus of basic and translation research in AD. Quantification and dissociation of the Aβ fractions in their soluble and insoluble forms, is a key factor in numerous AD studies.

New method: Here we provide a generalized sandwich-enzyme-linked-immuno-sorbent-assay (sELISA) protocol for quantification of human and murine Aβ1-40 and Aβ1-42 and dissociation of these peptides to their soluble-oligomeric and insoluble-fibrillar forms.

Results: We have validated the levels of soluble and insoluble Aβ1-40 and Aβ1-42 in the 5XFAD AD and the Ts65Dn Down-Syndrome (DS) mouse models in both the cortex, hippocampus and blood as follows: (1) blood levels of Aβ1-40 and Aβ1-42 are elevated in both mouse strains. (2) 5XFAD mice exhibit elevated soluble and insoluble Aβ1-40 in cortical and hippocampal tissues, soluble Aβ1-42 in the hippocampus, and insoluble Aβ1-42in both cortical and hippocampal tissues (3) Ts65Dn mice exhibit elevated levels of Aβ1-40 in the cortex.

Comparison with existing methods: Several methodologies have been proposed for the high throughput measure of Aβ, including HPLC-mass-spectrometry, micro-immunoelectrodes, immunoprecipitation and ELISA. Although commercial sELISA kits are widely used, herein, we describe a more accessible and cost-effective in-house protocol enabling to measure either human or murine, soluble and insoluble Aβ1-40 and Aβ1-42 levels.

Conclusions: We provide a streamlined and accessible protocol for the assessment of soluble and insoluble Aβ1-40 and Aβ1-42 levels from mouse or human origins, enabling a higher accessibility for researchers in the field to generate reliable Aβ-related measurements.

Keywords: 5XFAD; Alzheimer’s disease; Amyloid-beta; Aβ; ELISA; Ts65Dn.

MeSH terms

  • Amyloid beta-Peptides / analysis*
  • Amyloid beta-Peptides / metabolism
  • Animals
  • Blood Chemical Analysis / methods
  • Brain Chemistry
  • Cerebral Cortex / metabolism
  • Disease Models, Animal
  • Down Syndrome / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Hippocampus / metabolism
  • Humans
  • Male
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Peptide Fragments / analysis*
  • Peptide Fragments / metabolism
  • Reproducibility of Results

Substances

  • Amyloid beta-Peptides
  • Peptide Fragments
  • amyloid beta-protein (1-40)
  • amyloid beta-protein (1-42)