Matrix metalloproteinases (MMPs) belong to a family of secreted or membrane-associated zinc
endopeptidases capable of digesting extracellular matrix components. MMP-2 (72-kDa gelatinase-A/type IV collagenase) is responsible for degradation of collagen, fibronectin, laminin, and elastin. It has been shown that MMP-2 plays a key role in angiogenesis, tumor cell invasion, and metastasis. MMP-2 is also considered a therapeutic target for cancer.
The SensoLyte® Plus 520 MMP-2 Assay Kit is designed specifically for detecting MMP-2 activity in biological samples, such as culture medium, serum, plasma, synovial fluid, and tissue homogenate, which may contain multiple MMPs. Members of the MMP family have poor substrate
sequence specificity, thus using peptide substrate alone to differentiate the activity of a particular MMP from other MMPs is inadequate. A monoclonal anti-human-MMP-2 antibody is therefore employed to pull down MMP-2 from the mixture. MMP-2 activity is then quantified by a 5-FAM/QXL™ 520 fluorescence resonance energy transfer (FRET) peptide (Scheme 1). The long wavelength fluorescence of 5-FAM is less interfered by the autofluorescence of components in biological samples and test compounds. The assay can detect as low as 3 ng/mL active MMP-2 enzyme. The assay has no cross-reactivity with human MMP-1, 3, 7, 8, 9, 10, 12, 13, 15, 16 and has minimal cross-reactivity with MMP-14).