520 MMP FRET Substrate 3

Product Name: 520 MMP FRET Substrate 3

Sequence One Letter Code: QXL® 520-PLGC(Me)HAr-K(5-FAM)-NH2

Sequence Three Letter Code: QXL® 520-Pro-Leu-Gly-Cys(Me)-His-Ala-D-Arg-Lys(5-FAM)-NH2

Molecular Weight: 1743.9

Purity: 95%

Form: Lyophilized

Storage Conditions: - 20 °C Protected from light

Research Area: Cancer Disease Research

Conjugation: Conjugated

Conjugation Type: Double dyes

Code Nacres: NA.26

Application: 520 MMP FRET Substrate 3 is a quenched fluorogenic peptide engineered for sensitive detection of matrix metalloproteinase activity across multiple isoforms, including MMP-1, -2, -8, -9, -12, -13, and -14. Proteolytic cleavage separates the donor and quencher components, generating a fluorescence signal at 494/521 nm suitable for real-time kinetic monitoring. MMPs are zinc-dependent endopeptidases that orchestrate extracellular matrix degradation and regulate bioactive molecule processing in pathological and physiological contexts. This substrate enables quantitative enzyme characterization, inhibitor evaluation, and comparative activity profiling in both purified enzyme systems and cell-based assays. It is broadly applied in cancer invasion research, inflammation studies, angiogenesis modeling, and tissue remodeling investigations where accurate measurement of protease activity is required.

Current Research: 520 MMP FRET Substrate 3 is a quenched fluorogenic peptide developed for sensitive and quantitative detection of matrix metalloproteinase (MMP) activity across multiple isoforms, including MMP-1, -2, -8, -9, -12, -13, and -14. The substrate incorporates a fluorescence resonance energy transfer (FRET) donor–quencher pair that suppresses fluorescence in the intact peptide. Upon proteolytic cleavage by active MMPs, the donor fluorophore is separated from the quencher, producing a measurable fluorescence increase at Ex/Em 494/521 nm. This design enables continuous, real-time monitoring of protease activity in both biochemical and cell-based systems. Matrix Metalloproteinases in Physiology and Disease MMPs are zinc-dependent endopeptidases responsible for degradation and remodeling of extracellular matrix (ECM) components such as collagen, elastin, fibronectin, and laminin. Beyond structural matrix turnover, MMPs regulate the processing of cytokines, chemokines, and growth factors, thereby influencing cell migration, angiogenesis, inflammation, and tissue repair. Dysregulated MMP activity is implicated in a range of pathological conditions, including tumor invasion and metastasis, chronic inflammatory disorders, cardiovascular remodeling, fibrosis, and neurodegenerative diseases. Quantitative and isoform-inclusive measurement of MMP activity is therefore essential for mechanistic research and therapeutic development. FRET-Based Detection Mechanism The FRET architecture of 520 MMP FRET Substrate 3 allows homogeneous, separation-free detection of enzymatic cleavage. In the uncleaved state, energy transfer between the donor and quencher minimizes fluorescence emission. Proteolytic cleavage disrupts this interaction, generating a fluorescence signal proportional to enzymatic turnover. This configuration supports: Real-time kinetic analysis Determination of reaction velocity Quantitative calculation of kinetic parameters (K_m, V_max) Sensitive detection at low enzyme concentrations The excitation/emission profile (494/521 nm) aligns with commonly available fluorescence microplate readers, facilitating integration into high-throughput workflows. Broad Isoform Coverage This substrate is recognized by several key MMP isoforms, including collagenases (MMP-1, MMP-8, MMP-13), gelatinases (MMP-2, MMP-9), macrophage elastase (MMP-12), and membrane-type MMP-14 (MT1-MMP). Its broad reactivity makes it particularly useful for comparative activity profiling and for assessing proteolytic contributions in complex biological samples where multiple MMPs may be active. Applications in Cancer and Angiogenesis Research In oncology, MMP-mediated ECM degradation facilitates tumor cell invasion, basement membrane disruption, and metastatic dissemination. 520 MMP FRET Substrate 3 is commonly used to quantify protease activity in tumor cell-conditioned media, invasion assays, and 3D matrix remodeling models. In angiogenesis studies, MMP activity enables endothelial cell migration and vessel sprouting by remodeling perivascular matrix structures. Real-time monitoring of proteolysis supports investigation of pro-angiogenic signaling and evaluation of anti-angiogenic therapeutic candidates. Inflammation and Tissue Remodeling MMPs contribute to inflammatory signaling through cytokine processing and leukocyte recruitment. This substrate supports measurement of protease activity in macrophage, neutrophil, and fibroblast cultures under inflammatory stimulation. In fibrosis and wound-healing research, quantitative MMP assessment provides insight into matrix turnover and tissue repair dynamics. Inhibitor Screening and Enzyme Profiling 520 MMP FRET Substrate 3 is suitable for: High-throughput inhibitor screening Structure–activity relationship (SAR) studies Comparative analysis of MMP inhibitor selectivity Evaluation of recombinant enzyme activity Its robust fluorescence response and reproducibility ensure reliable data acquisition across multiple assay formats. Experimental Advantages Broad recognition across major MMP isoforms Continuous fluorescence-based readout High signal-to-background ratio Compatible with purified enzyme and cell-based assays Adaptable to microplate-based high-throughput platforms Overall, 520 MMP FRET Substrate 3 provides a sensitive and versatile platform for investigating matrix metalloproteinase activity. By enabling accurate, real-time quantification of proteolysis, it supports mechanistic studies of extracellular matrix remodeling, tumor invasion, inflammatory signaling, and therapeutic inhibitor development in translational research settings.