Cathepsins L and S activity detection is not as sensitive, most likely due to cathepsin K being a much more powerful collagenase, but here, we have expanded its utility and demonstrated its multiplex capacity to detect cathepsins K, L, and S in cell or tissue preparations from breast, lung, and cervical tumors to profile cathepsin activity at increasing stages of cancer progression and provide a new tool to screen pathological specimens for previously undetectable cathepsin activity. Here, CTSL is linked to cancer.