Cold Spring Harb Protoc. 2020 Jul 1;2020(7):099242. doi: 10.1101/pdb.top099242.
This introduction outlines general strategies for labeling proteins, with an emphasis on methods that are used primarily for labeling antibodies. It covers the specific site of modification, cross-linker options, types of labels, and postlabeling cleanup methodology, along with the advantages and disadvantages of each method. In general, polyclonal antibodies are more versatile and resistant to activity loss than are monoclonal antibodies. Greater care must be taken when labeling monoclonal antibodies to ensure a quality conjugate. The methods outlined here can be adapted for a variety of labels including multiple labels on the same immunoglobulin. The most important consideration when undertaking an antibody labeling experiment is to maintain the activity of the antibody. This is an empirical process and will often require additional experiments to optimize the label of a particular antibody. When successful, these reagents are very useful and adaptable biomolecules. This introduction provides the reader with methods and options for producing a variety of labeled immunological tools.
本文概述了用于标记蛋白质的一般策略，重点介绍了主要用于标记抗体的方法。它涵盖了修饰的特定部位、交联剂选择、标记类型以及标记后清洗方法，以及每种方法的优缺点。一般来说，多克隆抗体比单克隆抗体更通用且不易失去活性。在标记单克隆抗体时，必须更加小心，以确保获得高质量的缀合物。本文中概述的方法可以适用于多种标记物，包括同一免疫球蛋白上的多个标记物。进行抗体标记实验时最重要的考虑因素是保持抗体的活性。这是一个经验过程，通常需要进行额外的实验来优化特定抗体的标记。当成功时，这些试剂是非常有用和适应性强的生物分子。本文为读者提供了生产各种标记免疫工具的方法和选择。
