Antibody Elution Method for Multiple Immunohistochemistry on Primary Antibodies Raised in the Same Species and of the Same Subtype
Antibody Elution Method for Multiple Immunohistochemistry on Primary Antibodies Raised in the Same Species and of the Same Subtype
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DOI：
10.1369/jhc.2009.953240
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发表时间：
2009-06-01
影响因子：
3.2
通讯作者：
Stanescu, Radu
中科院分区：
文献类型：
--
作者：
Pirici, Daniel;Mogoanta, Laurentiu;Stanescu, Radu
IHC中的双重或多重抗原标记通常依赖于在不同物种中产生的一抗或不同IgG同种型的存在，以确保二次检测系统的特异性标记。然而，合适的一抗对并不总是可用或最佳选择（例如，诊断工具）。在过去的几年中，已经提出了几种方法来克服这一点，但它们都没有提供可靠的双重或多重酶或荧光IHC所需的灵活性。我们在这里提出了一个程序，洗脱抗体后的第一轮免疫标记，结合沉淀为基础的检测系统，允许多个IHC轮，即使是在同一物种和IgG同种型的一抗。与其他方法相比，该方法确保了可靠的酶或荧光染色，没有交叉反应性，也没有组织抗原性的损失，从而为共定位研究和病理诊断提供了一个灵活的工具。本手稿包含在线补充材料，请访问http://www.jhc.org在线查看这些材料。（J Histochem Cytochem 57：567-575，2009）
Double or multiple antigen labeling in IHC classically relies on the existence of primary antibodies raised in different species or of different IgG isotypes to ensure the specific labeling with the secondary detection systems. However, suitable pairs of primary antibodies are not always available or the best choice (e.g., as diagnostic tools). During the last few years, several methods have been proposed to overcome this, but none of them offers the flexibility needed for reliable double or multiple enzymatic or fluorescent IHC. We present here a procedure that elutes the antibodies after a first round of immunolabeling, which, in combination with precipitation-based detection systems, allows multiple IHC rounds even for primary antibodies raised in the same species and IgG isotype. Compared with other proposed methods, this procedure ensures a reliable enzymatic or fluorescent staining without cross-reactivity and without loss of tissue antigenicity, thus offering a flexible tool for colocalization studies and pathological diagnosis. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. (J Histochem Cytochem 57:567-575, 2009)