Double immunofluorescent staining of rat macrophages in formalin-fixed paraffin-embedded tissue using two monoclonal mouse antibodies.
Double immunofluorescent staining of rat macrophages in formalin-fixed paraffin-embedded tissue using two monoclonal mouse antibodies.
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使用两种单克隆小鼠抗体，在福尔马林固定石蜡包裹的组织中对大鼠巨噬细胞的双重免疫荧光染色。
DOI：
10.1007/s00418-015-1364-9
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发表时间：
2015-12
影响因子：
2.3
通讯作者：
Appleyard CB
中科院分区：
文献类型：
--
作者：
Isidro RA;Isidro AA;Cruz ML;Hernandez S;Appleyard CB
由于抗体可用性的限制，使用来自两种不同宿主物种的抗体对组织或细胞中的一种以上蛋白质进行双重免疫染色的常规方法并不总是可行的。先前报道的用于用源自相同物种的抗体对相同组织或细胞进行多重免疫染色的方法在其复杂性、灵敏度和防止抗体之间的不希望的相互作用的方法方面是不同的。在不断扩大的巨噬细胞生物学领域中，对小鼠和人类巨噬细胞的了解比大鼠巨噬细胞要多得多。来自不同物种的经验证和充分表征的单克隆抗体的有限可用性是阻碍大鼠巨噬细胞生物学进步的一个因素。在这里，我们描述了一种免疫染色方法，用于识别和检查大鼠巨噬细胞，是足够敏感的福尔马林固定石蜡包埋的组织中使用，只使用市售的试剂和抗体。该方法可用于帮助表征大鼠巨噬细胞中的生理学和病理生理学过程，并且可适于与来自相同物种来源的任何两种抗体一起使用，只要其中一种抗体是生物素化的。
The conventional approach of double immunostaining to visualize more than one protein in tissues or cells using antibodies from two different host species is not always feasible due to limitations with antibody availability. Previously reported methodologies for performing multiple immunostains on the same tissue or cells with antibodies originating from the same species are varied in their complexity, sensitivity, and approach to prevent unwanted interactions between antibodies. In the ever-expanding field of macrophage biology, much more is known about mouse and human macrophages than their rat counterparts. The limited availability of validated and well-characterized monoclonal antibodies from different species is one factor responsible for preventing advances in rat macrophage biology. Here we describe an immunostaining method for identifying and examining rat macrophages that is sufficiently sensitive for use in formalin-fixed paraffin embedded tissue and that uses only commercially available reagents and antibodies. This method can be used to help characterize both physiological and pathophysiological processes in rat macrophages, and can be adapted for use with any two antibodies from the same species of origin as long as one of the antibodies is biotinylated.