Characterization of a protein of interest during development is essential for functional studies. A general strategy for understanding the function of a particular protein involves the generation of null mutations, or treatment with drugs, that interfere with its activity. To demonstrate that the synthesis, stability, or activity of a protein has been affected, accurate and efficient detection of low amounts of protein is essential. This can be achieved by immunohistochemistry or by western blot. Here we describe a method for the detection of proteins from single de-yolked zebrafish embryos. This procedure includes a fixation step and the concomitant elimination of lipids from the yolk cell. We show that this approach allows the rapid analysis of proteins in embryos without having to manually remove the yolk. This method provides a convenient alternative for genotyping of mutant embryos as early as the 128 cell stage. In addition, in drug- or morpholino-treated embryos, the correlation between the penetrance of a phenotype and the concentration of a protein present may be established.

中文翻译：

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单个斑马鱼胚胎的蛋白质纯化和蛋白质印迹检测。

在开发过程中感兴趣的蛋白质的表征对于功能研究至关重要。理解特定蛋白质功能的一般策略涉及产生干扰其活性的无效突变或药物治疗。为了证明蛋白质的合成，稳定性或活性受到影响，准确有效地检测少量蛋白质至关重要。这可以通过免疫组织化学或蛋白质印迹来实现。在这里，我们描述了一种从单个脱黄斑马鱼胚胎中检测蛋白质的方法。该过程包括固定步骤和伴随的从卵黄细胞中清除脂质。我们表明，这种方法可以快速分析胚胎中的蛋白质，而无需手动去除蛋黄。该方法为早于128个细胞阶段的突变体胚胎的基因分型提供了一种方便的替代方法。另外，在药物或吗啉代处理的胚胎中，可以确定表型的渗透率与存在的蛋白质浓度之间的相关性。