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

STAR Protocols (Dec 2024)

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

Affiliations

Ying Lin: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
Shekhar Singh: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
Chong Xu: 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
Zeyu Wang: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
Cailin Feng: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
Dongyang Jiang: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
Lingfeng Luo: Division of Vascular Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford, CA, USA
Weiming Li: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
Wenliang Che: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China; Corresponding author
Guofu Zhu: Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China; Corresponding author

Journal volume & issue: Vol. 5, no. 4
p. 103519

Abstract

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Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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