Detection and Manipulation of the Stress Response Protein Metallothionein
Sadikshya Bhandari, Clare Melchiorre, Kristen Dostie, Debby Laukens, Lindsey Devisscher, Ariel Louwrier, Amy Thees, Michael A. Lynes
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引用次数: 12
Abstract
Metallothioneins (MTs) are small molecular weight stress response proteins that play a central role as reservoir of essential divalent heavy metal cations such as zinc and copper, and also can diminish the effects of toxic heavy metals such as mercury and cadmium. Historically, MT has been considered to be an intracellular protein with roles to play in the management of heavy metals, as a regulator of cellular redox potential, and as a buffer of free radicals. Our recent studies have highlighted immunomodulatory role of MT in inflammatory diseases and also in the progression of metastatic cell movement. Hence, manipulation and detection of MT is essential for its possible use as a diagnostic and in therapeutic interventions of chronic inflammation. This review describes procedures used to detect MT using techniques such as western immunoblot, competition ELISA, flow cytometry and immunohistochemistry. Additionally, it also describes the use of a colorimetric cell proliferation assay (CellTiter 96 AQueous One Solution/MTS) to study the proliferative effect of MT. © 2017 by John Wiley & Sons, Inc.
应激反应蛋白金属硫蛋白的检测与调控
金属硫蛋白(MTs)是一种小分子量的应激反应蛋白,作为锌和铜等必需二价重金属阳离子的储存库起着核心作用,也可以减轻汞和镉等有毒重金属的影响。从历史上看,MT被认为是一种细胞内蛋白,在重金属管理中发挥作用,作为细胞氧化还原电位的调节剂,并作为自由基的缓冲剂。我们最近的研究强调了MT在炎症性疾病和转移细胞运动进展中的免疫调节作用。因此,MT的操作和检测对于其作为慢性炎症的诊断和治疗干预的可能用途至关重要。本文综述了使用免疫印迹、竞争ELISA、流式细胞术和免疫组织化学等技术检测MT的方法。此外,它还描述了使用比色细胞增殖试验(CellTiter 96水溶液/MTS)来研究MT的增殖作用。©2017 by John Wiley &儿子,Inc。
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