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

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.