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Catalog#: 2260001
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Analyzing the Quenchable Iron Pool in Murine Macrophages by Flow Cytometry
[Abstract]  Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on cellular iron homeostasis in macrophages remains challenging due to the limited amount of available methods using radioactive 59Fe isotopes or strong iron chelators, which might be inapplicable in certain experimental settings. This protocol describes the analysis of the quenchable iron pool (QIP) in macrophages by loading these cells with exogenous iron-complexes. Thereby, the cytoplasmic iron pool can be determined, since the iron uptake ability of macrophages inversely correlates with intracellular iron levels. Thus, this assay ... [摘要]  [摘要 ] 驻留在组织中的巨噬细胞对于在细胞和全身水平上严格调节铁代谢至关重要,因为细微的铁改变会增加对微生物感染的敏感性或引发多种疾病。然而,由于使用放射性59 Fe同位素或强铁螯合剂的可用方法数量有限,因此巨噬细胞中细胞铁稳态的研究仍然具有挑战性,这在某些实验环境中可能不适用。该协议 描述了通过向这些细胞加载外源铁配合物来分析巨噬细胞中的可淬灭铁池(QIP)。因此,由于巨噬细胞的铁摄取能力与细胞内铁水平成反比,因此可以确定细胞质的铁库。因此,该测定法能够准确分析细胞质铁通量的微小变化,并且几乎适用于所有实验室环境。另外,该方案还可以用于体外和体内的其他免疫细胞类型。

[背景 ] 由于用于全身铁代谢它们的中心调节功能,在多种组织中的巨噬细胞中迅速地改变在不同的刺激的遭遇它们的细胞内的铁水平的微生物感染和疾病(Nairz 等人,2017) 。值得注意的是,大多数细胞中的铁离子被细胞质中的主要铁存储蛋白铁蛋白结合或定位于线粒体或溶酶体等区室(Ma 等,2015 ; Soares和Hamza,2016 ...