Rega Camilla, Pardo Mercedes, Martin Lesley-Ann, Choudhary Jyoti
The Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.
The Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.
STAR Protoc. 2025 Mar 21;6(1):103547. doi: 10.1016/j.xpro.2024.103547. Epub 2025 Jan 10.
Biotinylation by antibody recognition (BAR) is an antibody-based approach for mapping proximal protein interactions in cells. Here, we present a protocol to biotinylate and identify proximal proteins using BAR. We describe steps for defining proximity labeling reaction conditions, assessing enrichment using western blot, and sample preparation for mass spectroscopy analysis. We then detail procedures for data analysis and identifying proximal proteins. This approach differs from standard proximity labeling techniques, which rely on genetically engineered enzymes fused to the target protein. For complete details on the use and execution of this protocol, please refer to Rega et al..
通过抗体识别进行生物素化（BAR）是一种基于抗体的方法，用于绘制细胞中近端蛋白质相互作用图谱。在此，我们展示了一种使用BAR对近端蛋白质进行生物素化和鉴定的方案。我们描述了定义邻近标记反应条件、使用蛋白质印迹评估富集以及用于质谱分析的样品制备步骤。然后，我们详细介绍了数据分析和鉴定近端蛋白质的程序。这种方法不同于标准的邻近标记技术，后者依赖于与目标蛋白融合的基因工程酶。有关本方案使用和执行的完整详细信息，请参考Rega等人的研究。
