Use of phospholipase A2 for antigen retrieval in zebrafish whole-mount immunohistochemistry.
Use of phospholipase A2 for antigen retrieval in zebrafish whole-mount immunohistochemistry.
复制标题
使用磷脂酶 A2 在斑马鱼整体免疫组织化学中进行抗原修复。
DOI：
10.1089/zeb.2009.0588
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发表时间：
2009
期刊：
影响因子：
2
通讯作者：
Wallace,KennethN
中科院分区：
文献类型：
--
作者：
Akhtar,Tanveer;Li,Jiannan;Olden,Tasha;Wallace,KennethN
斑马鱼的一个优势是我们可以使用整体免疫组织化学来识别蛋白质表达的空间和时间模式。为了让一抗与其靶标相互作用，大多数组织必须进行某种类型的抗原修复。许多修复技术利用蛋白质消化酶来获取抗原。在这里，我们研究了使用磷脂酶 A2 (PLA2) 作为抗原修复的唯一酶以及与低浓度的蛋白酶 K 的组合。当用作唯一的酶时，与 PLA2 一起使用的蛋白酶 K 浓度无法暴露抗原。我们证明 PLA2 对核抗原和细胞质抗原都有用，但对细胞外基质成分无效。
An advantage in zebrafish is that we can identify spatial and temporal patterns of protein expression using whole-mount immunohistochemistry. To allow primary antibodies to interact with their targets, most tissues must undergo some type of antigen retrieval. Many retrieval techniques have utilized protein-digesting enzymes to access antigens. Here we investigate the use of phospholipase A2(PLA2) as the sole enzyme for antigen retrieval as well as in combination with low concentrations of proteinase K. Concentrations of proteinase K used with PLA2are unable to expose the antigen when used as the sole enzyme. We demonstrate that PLA2is useful for both nuclear and cytoplasmic antigens but not for extracellular matrix components.