At the annual conference of the American Society for Cell Biology, AnaSpec presented a technical poster documenting improved technology for MMP detection assays. In a previous poster, AnaSpec documented the use of highly sensitive FRET-peptide substrates
for MMP detection. The latest poster documented the incorporation of immunocapture technologies to achieve greatly improved specificity.
Matrix metalloproteinases (MMPs) belong to a family of secreted or membrane-associated zinc endopeptidases capable
of digesting extracellular matrix components. MMPs play roles in the multiple stages of tumor development and metastasis, rheumatoid arthritis, wound healing, angiogenesis, and other pathological and physiological events.
When a FRET-peptide based assay for MMPs is used in cell or tissue lysates, non-specific cleavages by MMPs and other proteases are also measured, which may be simultaneously present in biological fluids. Since multiple MMPs generally co-exist in biological samples, the assay employing the FRET peptide alone makes differentiation between the activities of different MMPs difficult, if not impossible. To solve this issue, MMP antibodies are used to first immobilize a specific MMP from biological samples. After washing, the FRET peptide is applied to measure the activity.
FRET assays that incorporate this immunocapture technology are highly sensitive and specifically detect the activity of a particular MMP in a mixed biological sample. The assay sensitivity can reach picogram amounts of enzyme with a large and linear assay range. The assay can also be used to study the interaction between different MMPs. Immunocapture-based FRET technology is the backbone of AnaSpec’s EnzoLyte PlusTM MMP Assay Kits.
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