An activatable fluorescent probe from indocyanine was developed for the detection of tumor-enriched γ-glutamyltranspeptidase (γGT). The probe exhibited a dramatic fluorescence enhancement (F/F0 = 10) as well as a bathochromic shift (>100 nm) upon the treatment of γGT with a low limit of detection of 0.15 unit/L and was further successfully applied as a sensitive probe for γGT in the mouse model of colon cancer.