520 MMP FRET Substrate 15

Product Name: 520 MMP FRET Substrate 15

Sequence One Letter Code: QXL® 520 -γ-Abu-PQGL-Dab(5-FAM)-AK-NH2

Sequence Three Letter Code: QXL® 520 -γ-Abu-Pro-Gln-Gly-Leu-Dab(5-FAM)-Ala-Lys-NH2

Molecular Weight: 1646.8

Purity: 95%

Form: Lyophilized

Storage Conditions: - 20 °C Protected from light

Research Area: peptide substrate

Conjugation: Conjugated

Conjugation Type: Double dyes

Code Nacres: NA.26

Application: 520 MMP FRET Substrate 15 is a fluorogenic peptide developed for broad detection of matrix metalloproteinase activity in extracellular matrix and tissue remodeling studies. It is efficiently cleaved by multiple MMPs, including MMP-1, -2, -7, -8, -12, -13, and -14. Proteolytic cleavage disrupts FRET quenching and generates fluorescence at Ex/Em 494/521 nm, enabling sensitive kinetic analysis, inhibitor evaluation, and comparative profiling of MMP function in biological samples.

Current Research: 520 MMP FRET Substrate 15 is a fluorogenic peptide substrate engineered for broad and sensitive detection of matrix metalloproteinase (MMP) activity in studies of extracellular matrix (ECM) remodeling and tissue pathology. The substrate incorporates a fluorescence resonance energy transfer (FRET) donor–quencher pair, maintaining low baseline fluorescence in its intact state. Upon enzymatic cleavage, quenching is disrupted, resulting in a measurable fluorescence signal at approximately Ex 494 nm / Em 521 nm. This substrate is efficiently hydrolyzed by multiple MMP family members, including MMP-1, -2, -7, -8, -12, -13, and -14, enabling comprehensive profiling of metalloproteinase activity in complex biological systems. Biological Context MMPs are zinc-dependent endopeptidases that regulate ECM turnover and influence key physiological and pathological processes, including: Collagen and elastin degradation Angiogenesis Wound healing Inflammatory responses Tumor invasion and metastasis Fibrosis and connective tissue disorders Because many MMPs exhibit overlapping substrate specificities, broad-spectrum substrates such as 520 MMP FRET Substrate 15 are valuable for measuring total or composite MMP activity in biological samples. Mechanism of Detection The substrate contains: An MMP-recognition peptide sequence A fluorophore emitting at ~521 nm A proximal quencher that suppresses fluorescence via FRET In the intact molecule, energy transfer from the donor to the quencher prevents fluorescence emission. Upon cleavage by active MMPs: The peptide backbone is hydrolyzed. Donor and quencher are spatially separated. Fluorescence intensity increases proportionally to protease activity. This design enables real-time kinetic monitoring with high sensitivity and minimal background interference. Research Applications 1. Broad MMP Activity Profiling The substrate supports quantification of total MMP activity in: Conditioned cell culture media Tissue homogenates Serum or plasma samples ECM-rich experimental systems 2. Tissue Remodeling and Fibrosis Research MMP-1, -2, -13, and -14 play prominent roles in collagen turnover and matrix degradation. The substrate facilitates functional analysis in models of fibrosis, arthritis, and connective tissue remodeling. 3. Angiogenesis and Tumor Biology MMP-mediated ECM degradation is essential for endothelial migration and tumor invasion. This substrate enables evaluation of proteolytic activity in cancer progression and angiogenic studies. 4. Inhibitor Screening The FRET-based fluorescence readout is compatible with high-throughput screening of MMP inhibitors. Decreased fluorescence relative to control indicates effective protease inhibition. Advantages Broad reactivity across major MMP subtypes High sensitivity and low background fluorescence Real-time kinetic measurement capability Suitable for complex biological samples Compatible with microplate fluorescence instrumentation Experimental Considerations Optimal assay conditions require appropriate buffer composition, including zinc and calcium ions essential for MMP catalytic activity. Activation of latent pro-MMPs (e.g., via APMA treatment) may be necessary prior to analysis. Inclusion of selective MMP inhibitors helps confirm assay specificity in mixed protease systems.