科研之友
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摘要
Background: Amyloid-beta (A beta), a hallmark of Alzheimer's disease (AD), has long been a focus of basic and translation research in AD. Quantification and dissociation of the A beta 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 A beta(1-40) and A beta(1-42) and dissociation of these peptides to their soluble-oligomeric and insoluble-fibrillar forms. Results: We have validated the levels of soluble and insoluble A beta(1-40) and A beta(1-42) 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 A beta(1-40) and A beta(1-42) are elevated in both mouse strains. (2) 5XFAD mice exhibit elevated soluble and insoluble A beta(1-40) in cortical and hippocampal tissues, soluble A beta(1-42) in the hippocampus, and insoluble A beta(1-42) both cortical and hippocampal tissues (3) Ts65Dn mice exhibit elevated levels of A beta(1-40) in the cortex. Comparison with existing methods: Several methodologies have been proposed for the high throughput measure of A beta, 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 A beta(1-40) and A beta(1-42) levels. Conclusions: We provide a streamlined and accessible protocol for the assessment of soluble and insoluble A beta(1-40)and A beta(1-42) levels from mouse or human origins, enabling a higher accessibility for researchers in the field to generate reliable A beta-related measurements.
- 出版日期2017-11-1
