Valle Maria Luisa, Getaneh Bitseat, Loveland James, Erdjument-Bromage Hediye, William Christopher, Neubert Thomas A, Rostagno Agueda, Ghiso Jorge
Department of Pathology, New York University Grossman School of Medicine, New York, NY 10016, USA.
Department of Pathology, New York University Grossman School of Medicine, New York, NY 10016, USA.
STAR Protoc. 2025 Jun 20;6(2):103864. doi: 10.1016/j.xpro.2025.103864. Epub 2025 Jun 2.
Alzheimer's disease (AD) is characterized by the deposition of full-length and truncated amyloid beta (Aβ) species within brain parenchyma and cerebral vessels. However, Aβ truncated species remain understudied. Here, we present a protocol for culture and characterization of a mouse monoclonal antibody targeting N-terminally truncated proteoforms starting at position 4. We describe a detailed procedure for hybridoma culture, antibody collection, and isolation via affinity chromatography. We then describe steps for target validation via dot blot, as well as potential applications. For complete details on the use and execution of this protocol, please refer to Cabrera et al. and Rostagno et al..
阿尔茨海默病（AD）的特征是全长和截短的β淀粉样蛋白（Aβ）在脑实质和脑血管内沉积。然而，截短的Aβ种类仍未得到充分研究。在此，我们介绍一种针对从第4位开始的N端截短蛋白亚型的小鼠单克隆抗体的培养和表征方案。我们描述了杂交瘤培养、抗体收集以及通过亲和层析进行分离的详细步骤。然后我们描述了通过斑点印迹进行靶点验证的步骤以及潜在应用。有关本方案使用和执行的完整详细信息，请参考卡布雷拉等人以及罗斯塔尼奥等人的研究。
