Forsthoefel David J, Ross Kelly G, Newmark Phillip A, Zayas Ricardo M
Functional and Chemical Genomics Research Program, Oklahoma Medical Research Foundation, Oklahoma, OK, USA.
Department of Biology, San Diego State University, San Diego, CA, USA.
Methods Mol Biol. 2018;1774:353-366. doi: 10.1007/978-1-4939-7802-1_10.
Efforts to elucidate mechanisms of regeneration in the planarian Schmidtea mediterranea have included the application of immunocytochemical methods to detect specific molecules and label cells and tissues in situ. Here we describe methods for immunofluorescent labeling of whole mount planarians. We outline protocols for fixation and steps for processing animals prior to immunolabeling, incorporating commonly utilized reagents for mucus removal, pigment bleaching, tissue permeabilization, and antigen retrieval. Because processing steps can mask or degrade antigens, we also recommend protocol parameters that can be tested simultaneously to optimize sample preparation for novel antibodies.
为阐明地中海涡虫再生机制所做的努力，包括应用免疫细胞化学方法来检测特定分子并原位标记细胞和组织。在此，我们描述了对整装涡虫进行免疫荧光标记的方法。我们概述了固定方案以及免疫标记前处理动物的步骤，其中纳入了常用于去除黏液、漂白色素、通透组织以及抗原修复的试剂。由于处理步骤可能会掩盖或降解抗原，我们还推荐了可同时进行测试的方案参数，以优化针对新型抗体的样品制备。
