Fig. 3: Schematic of the multistep click-linking workflow. | Nature Communications

Fig. 3: Schematic of the multistep click-linking workflow.

From: Click-linking: a cell-compatible protein crosslinking method based on click chemistry

Fig. 3: Schematic of the multistep click-linking workflow.

Cells are first stabilized with high-concentration formaldehyde and then permeabilized to allow for the simultaneous introduction of two reactive click precursors. After bioconjugation and washing, CuAAC catalysts are added to complete the crosslink. After capping any remaining free azides, cells are harvested for sample workup. Reagent 1: 2,5-dioxopyrrolidin-1-yl 3-(prop-2-ynyloxy)propanoate. Reagent 2: and (2,5-dioxopyrrolidin-1-yl) 6-azidohexanoate. Reagent 3: 11,12-didehydro-Îł-oxodibenz[b,f]azocine-5(6H)-butanoic acid. Created in BioRender. Schriemer, D. (2025) https://BioRender.com/p82h674.

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