hSUMO1-AMC is prepared via the conjugation of 7-amino-4-methylcoumarin (AMC) to the C-terminal Gly of hSUMO1. This conjugation quenches the intrinsic fluorescence of AMC. Upon incubation with a protease recognizing hSUMOs, such as Senp1 or Senp2, AMC is released and the increase in fluorescence at 460 nm (Exc. 380 nm) can be measured. The enhanced purity of hSUMO1-AMC compared to that of other commercial vendors leads to a greater signal to noise ratio. This protein contains no extraneous tags.