Schmidt Stephen D, Mazzella Matthew J, Nixon Ralph A, Mathews Paul M
Center for Dementia Research, Nathan S. Kline Institute for Psychiatric Research, Orangeburg, NY, USA.
Methods Mol Biol. 2012;849:507-27. doi: 10.1007/978-1-61779-551-0_34.
The neuritic plaque in the brain of Alzheimer's disease patients consists of an amyloid composed primarily of Aβ, an approximately 4-kDa peptide derived from the amyloid precursor protein. Multiple lines of evidence suggest that Aβ plays a key role in the pathogenesis of the disease, and potential treatments that target Aβ production and/or Aβ accumulation in the brain as β-amyloid are being aggressively pursued. Methods to quantitate the Aβ peptide are, therefore, invaluable to most studies aimed at a better understanding of the molecular etiology of the disease and in assessing potential therapeutics. Although other techniques have been used to measure Aβ in the brains of AD patients and β-amyloid-depositing transgenic mice, the enzyme-linked immunosorbent assay (ELISA) is one of the most commonly used, reliable, and sensitive methods for quantitating the Aβ peptide. Here we describe methods for the recovery of both soluble and deposited Aβ from brain tissue and the subsequent quantitation of the peptide by sandwich ELISA.
阿尔茨海默病患者大脑中的神经炎性斑块由主要由Aβ组成的淀粉样蛋白构成，Aβ是一种源自淀粉样前体蛋白的约4 kDa肽。多条证据表明，Aβ在该疾病的发病机制中起关键作用，目前正在积极探索以Aβ产生和/或大脑中作为β淀粉样蛋白的Aβ积累为靶点的潜在治疗方法。因此，定量Aβ肽的方法对于大多数旨在更好地理解该疾病分子病因以及评估潜在治疗方法的研究来说非常宝贵。尽管已使用其他技术来测量AD患者大脑和β淀粉样蛋白沉积转基因小鼠大脑中的Aβ，但酶联免疫吸附测定（ELISA）是定量Aβ肽最常用、可靠且灵敏的方法之一。在此，我们描述了从脑组织中回收可溶性和沉积性Aβ以及随后通过夹心ELISA对该肽进行定量的方法。
