Protocol for tyramide signal amplification immunohistochemical detection of Notch1 signaling in the vascular system

Ying Lin; Shekhar Singh; Chong Xu; Zeyu Wang; Cailin Feng; Dongyang Jiang; Lingfeng Luo; Weiming Li; Wenliang Che; Guofu Zhu

doi:10.1016/j.xpro.2024.103519

Protocol for tyramide signal amplification immunohistochemical detection of Notch1 signaling in the vascular system

STAR Protoc. 2024 Dec 10;5(4):103519. doi: 10.1016/j.xpro.2024.103519. Online ahead of print.

Authors

Ying Lin¹, Shekhar Singh¹, Chong Xu², Zeyu Wang¹, Cailin Feng¹, Dongyang Jiang¹, Lingfeng Luo³, Weiming Li¹, Wenliang Che⁴, Guofu Zhu⁵

Affiliations

¹ Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China.
² Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China; Department of Cardiology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi Medical Center, Nanjing Medical University, Wuxi, China.
³ Division of Vascular Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford, CA, USA.
⁴ Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China. Electronic address: chewenliang@tongji.edu.cn.
⁵ Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China. Electronic address: guofu_zhu@tongji.edu.cn.

PMID: 39661510
DOI: 10.1016/j.xpro.2024.103519

Abstract

Notch signaling is a pivotal regulator in the vascular system that is essential for development, angiogenesis, and maintaining vascular homeostasis. Here, we present a protocol for tyramide signal amplification (TSA) immunohistochemistry, tailored explicitly for detecting Notch signaling components in vascular tissues. We describe steps for utilizing tailored antigen retrieval techniques, specific blocking solutions, and a complex of avidin/biotin-horseradish peroxidase conjugate with tyramide, along with optimized washing steps. For complete details on the use and execution of this protocol, please refer to Zhu et al.¹.

Keywords: cell biology; developmental biology; model organisms; molecular biology.