Assays for matrix metalloprotein activity

To detect activities of MMP-1, MMP-2, MMP-3, MMP-7 and MMP-9, use radio-labeled fibrillar collagen (for MMP-1), gelatin (for MMP-2 and MMP-9) and carboxymethylated transferrin (for MMP-3 and MMP-7) substrates for the assays (1), although these can be substituted with commercially available FITC-labeled substrates and fluorogenic synthetic peptides for the MMPs. Zymography using gelatin or casein substrate gels is also useful to monitor gelatinolytic activity of MMP-2 and MMP-9 or caseinolytic activity of MMP-3 and MMP-7 in each column fraction, although zymography is not suitable for calculation of purification fold and recovery of proteinases.