Institute of Human Genetics, University of Göttingen Medical Center, Georg-August-Universität, Göttingen, Germany.
Department of Developmental Biology, Johann-Friedrich-Blumenbach Institute of Zoology and Anthropology, Göttingen Center of Molecular Biosciences, Göttingen, Germany.
Methods Mol Biol. 2021;2218:245-252. doi: 10.1007/978-1-0716-0970-5_19.
Immunohistochemistry has been widely used as a robust technique to determine the cellular and subcellular localization of proteins. This information ultimately helps to understand the function of these proteins and how biological processes are regulated. Antibodies applicable for labeling in zebrafish are limited, making immuno-staining challenging. Recently glyoxal fixation was rediscovered in tissue culture, mouse, rat, and Drosophila, expanding the list of effective antibodies for these species. Here, we compare a protocol for zebrafish staining using glyoxal as a fixative agent with PFA. We demonstrate that glyoxal fixation improves the antigenicity of some epitopes thereby increasing the number of useful antibodies in zebrafish.
免疫组织化学已被广泛用作确定蛋白质细胞和亚细胞定位的强大技术。这些信息最终有助于了解这些蛋白质的功能以及生物过程是如何被调控的。适用于斑马鱼标记的抗体有限，这使得免疫染色具有挑战性。最近，在组织培养、小鼠、大鼠和果蝇中重新发现了戊二醛固定，这扩大了这些物种中有效抗体的范围。在这里，我们比较了使用戊二醛作为固定剂的斑马鱼染色方案与 PFA。我们证明，戊二醛固定可以提高一些表位的抗原性，从而增加斑马鱼中有用抗体的数量。
