Antibody staining of Parhyale hawaiensis embryos.
Antibody staining of Parhyale hawaiensis embryos.
复制标题
Parhyale hawaiensis 胚胎的抗体染色。
作者：
E. J. Rehm;Roberta L Hannibal;R. C. Chaw;Mario A. Vargas;N. Patel
节肢动物身体计划的巨大多样性，加上我们对果蝇发育的详细了解，使节肢动物成为研究发育模式进化多样性和现存生命多样性的首要分类单元。特别是甲壳类动物，显示出一个显着的形态范围，并提供了一个有用的外类群的昆虫。片足类甲壳动物Parhyale hawaiensis正在成为节肢动物发育研究的模式生物。该方案提供了用于夏威夷假单胞菌胚胎的抗体染色的简化方案。该方法也适用于其他节肢动物和门。将固定的胚胎再水化，洗涤，用正常山羊血清封闭，并与一抗孵育过夜。然后清洗胚胎，并与过氧化物酶偶联的第二抗体一起孵育，所述第二抗体与第一抗体结合。随后的组织化学反应在抗体定位的那些细胞中产生黑色染色。
The great diversity of arthropod body plans, together with our detailed understanding of fruit fly development, makes arthropods a premier taxon for examining the evolutionary diversification of developmental patterns and hence the diversity of extant life. Crustaceans, in particular, show a remarkable range of morphologies and provide a useful outgroup to the insects. The amphipod crustacean Parhyale hawaiensis is becoming established as a model organism for developmental studies within the arthropods. This protocol provides a simplified protocol for antibody staining of P. hawaiensis embryos. The method also works well for other arthropods and phyla. Fixed embryos are rehydrated, washed, blocked with normal goat serum, and incubated overnight with primary antibody. Embryos are then washed and incubated with a peroxidase-conjugated secondary antibody that binds to the primary antibody. A subsequent histochemical reaction produces a black stain in those cells where antibodies have localized.