Work

Protocol for trapping transient endogenous formaldehyde in live cells to visualize its signaling

Year: 2025

Type: article

Abstract: Formaldehyde is emerging as a key signaling modulator. Here, we present Trami, a protocol that traps endogenous formaldehyde in live cells and converts its transient signals into stable proximal protein labels for subsequent signaling study. We describe probe design, live-cell administration, and imaging procedures for signaling deconvolution. Trami offers a powerful tool for elucidating formaldehyde-mediated signaling pathways. For complete details on the use and execution of this protocol, please refer to Pan et al. 1 • Details on steps for staining cells with the Trami probe • Procedures for immunofluorescence staining • Guidance on multiplex imaging • Instructions for correlating FA levels with proinflammatory signaling Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Formaldehyde is emerging as a key signaling modulator. Here, we present Trami, a protocol that traps endogenous formaldehyde in live cells and converts its transient signals into stable proximal protein labels for subsequent signaling study. We describe probe design, live-cell administration, and imaging procedures for signaling deconvolution. Trami offers a powerful tool for elucidating formaldehyde-mediated signaling pathways.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104108

Authors: Yuan Pan, Xing-Guang Liang, Xin Li

Institutions: Zhejiang University, First Affiliated Hospital Zhejiang University

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: National Natural Science Foundation of China, Natural Science Foundation of Zhejiang Province