520 MMP FRET Substrate 12

Product Name: 520 MMP FRET Substrate 12

Sequence One Letter Code: 5-FAM-RPKPYA-Nva-WM-K(QXL® 520)-NH2

Sequence Three Letter Code: 5-FAM-Arg-Pro-Lys-Pro-Tyr-Ala-Nva-Trp-Met-Lys(QXL® 520)-NH2

Molecular Weight: 2125.4

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 12 is a quenched fluorogenic peptide formulated for quantitative assessment of matrix metalloproteinase activity in vitro and in cell-based systems. Cleavage by MMP isoforms—including MMP-1, -2, -7, -8, -12, and -13—disrupts the FRET pair and produces a fluorescence signal at 494/521 nm. MMPs regulate extracellular matrix turnover and proteolytic processing of signaling mediators implicated in tumor progression, inflammation, and angiogenesis. This substrate enables kinetic enzyme profiling, inhibitor screening, and comparative isoform analysis under controlled conditions. Its robust fluorescence response and compatibility with standard fluorescence plate readers make it suitable for high-throughput enzymology workflows and translational research applications focused on tissue remodeling and disease-associated protease activity.

Current Research: 520 MMP FRET Substrate 12 is a quenched fluorogenic peptide designed for sensitive and quantitative detection of matrix metalloproteinase (MMP) activity in biochemical and cell-based assays. Built on a fluorescence resonance energy transfer (FRET) platform, the intact substrate exhibits minimal fluorescence due to close proximity between the donor fluorophore and quencher. Upon proteolytic cleavage by target MMP isoforms—including MMP-1, -2, -7, -8, -12, and -13—the donor and quencher are separated, resulting in a measurable fluorescence increase at Ex/Em 494/521 nm. This fluorescence change directly reflects enzymatic activity and supports real-time kinetic analysis. Role of MMPs in Biology and Disease Matrix metalloproteinases are zinc-dependent endopeptidases that mediate extracellular matrix (ECM) degradation and remodeling. They regulate structural components such as collagen, elastin, and gelatin, while also processing growth factors, cytokines, and chemokines. Through these functions, MMPs influence cell migration, angiogenesis, inflammation, and tissue repair. Aberrant MMP activity is strongly associated with pathological conditions, including tumor invasion and metastasis, chronic inflammatory diseases, fibrosis, cardiovascular remodeling, and neurodegenerative disorders. Quantitative assessment of MMP activity is therefore essential for mechanistic research and therapeutic development. FRET-Based Detection Mechanism The FRET design of 520 MMP FRET Substrate 12 enables continuous, homogeneous measurement of proteolytic activity without the need for secondary detection reagents. In the uncleaved state, fluorescence emission is suppressed due to energy transfer between the donor and quencher. Protease-mediated cleavage disrupts this interaction, generating a proportional fluorescence signal. Key advantages of this detection system include: Real-time kinetic monitoring High signal-to-background ratio Quantitative measurement of reaction velocity Compatibility with standard fluorescence instrumentation The excitation and emission wavelengths (494/521 nm) align with commonly available microplate readers, facilitating seamless integration into established laboratory workflows. Applications in Enzymology and Drug Discovery 520 MMP FRET Substrate 12 supports comprehensive enzymatic profiling, including: Determination of kinetic parameters (K_m, V_max, catalytic efficiency) Comparative activity analysis across MMP isoforms Evaluation of substrate specificity Assessment of enzyme activation and inhibition In drug discovery settings, the substrate is widely used for high-throughput screening (HTS) of MMP inhibitors. Because fluorescence generation is directly proportional to proteolytic cleavage, inhibitor potency can be accurately quantified under controlled conditions. This makes the substrate valuable for structure–activity relationship (SAR) studies and early-stage therapeutic validation. Cancer, Inflammation, and Angiogenesis Research MMP-mediated ECM degradation facilitates tumor cell invasion and metastatic dissemination. Monitoring MMP activity in tumor cell supernatants or co-culture systems provides insight into invasive phenotypes and tumor microenvironment remodeling. The substrate is also applied in 3D matrix degradation assays and spheroid invasion models. In inflammatory research, MMPs regulate leukocyte migration and cytokine processing. Dynamic measurement of protease activity in macrophage, neutrophil, or fibroblast cultures enables investigation of inflammatory signaling networks and tissue injury mechanisms. Similarly, in angiogenesis studies, MMP-driven matrix remodeling is essential for endothelial sprouting and vessel formation, making quantitative protease monitoring a critical component of vascular biology research. Experimental Advantages Broad reactivity with key MMP isoforms Continuous fluorescence-based readout High sensitivity and reproducibility Adaptable to in vitro and cell-based assays Suitable for high-throughput plate formats Because the assay is homogeneous and requires no separation steps, it reduces workflow complexity and variability. Its robust fluorescence response supports precise enzyme characterization across diverse research applications. Research Impact By enabling accurate, real-time measurement of MMP activity, 520 MMP FRET Substrate 12 provides a powerful tool for investigating extracellular matrix remodeling, protease-driven signaling, and disease-associated enzymatic dysregulation. Its compatibility with high-throughput enzymology platforms makes it particularly valuable in translational research focused on cancer progression, inflammatory disorders, and therapeutic inhibitor development.