Here, we first report a rapid and highly selective biocompatible ligation that proceeds via a strain-promoted prenyl-involved [2, 3]-Ene rearrangement process. We demonstrate the usefulness of naturally occurring strain-promoted β-caryophyllene with triazoline (PTAD)/Selectfluor in the study of tagging molecule-of-interest. Experiments in peptide (Histone H3 (1-21) and Myhc (614-629)) and protein (BSA, βLG, and HSP40) models exemplified the utility of the Ene-ligation for in vivo imaging and tracking.