520 MMP FRET Substrate 14

Product Name: 520 MMP FRET Substrate 14

Sequence One Letter Code: QXL® 520 -γ-Abu-P-Cha-Abu-Smc-HA-Dab(5-FAM)-AK-NH2 (Smc=S-Methyl-L-cysteine)

Sequence Three Letter Code: QXL® 520 -γ-Abu-Pro-Cha-Abu-Smc-His-Ala-Dab(5-FAM)-Ala-Lys-NH2 (Smc=S-Methyl-L-cysteine)

Molecular Weight: 1912.1

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 14 is a fluorogenic peptide substrate designed for comprehensive assessment of matrix metalloproteinase activity. It is cleaved by a wide spectrum of MMPs, including MMP-1, -2, -3, -7, -8, -9, -12, and -13, supporting broad enzymatic profiling. Cleavage relieves fluorescence quenching and produces a robust signal measurable at Ex/Em 494/521 nm. This substrate is suitable for kinetic measurements, inhibitor screening, and mechanistic studies of extracellular matrix remodeling and inflammatory signaling.

Current Research: 520 MMP FRET Substrate 14 is a fluorogenic peptide substrate engineered for broad and sensitive detection of matrix metalloproteinase (MMP) activity in extracellular matrix (ECM) research. The substrate incorporates a fluorescence resonance energy transfer (FRET) donor–quencher pair that maintains low baseline fluorescence in its intact form. Proteolytic cleavage by active MMPs disrupts quenching, generating a measurable fluorescence signal at approximately Ex 494 nm / Em 521 nm. This substrate is efficiently cleaved by a wide spectrum of MMP family members, including MMP-1, -2, -3, -7, -8, -9, -12, and -13, enabling comprehensive enzymatic profiling in diverse biological contexts. Biological Context Matrix metalloproteinases are zinc-dependent endopeptidases responsible for controlled degradation and remodeling of the extracellular matrix. They regulate key processes such as: Collagen and elastin turnover Cell migration and invasion Angiogenesis Cytokine and growth factor activation Inflammatory tissue remodeling Because multiple MMPs often act concurrently in physiological and pathological settings, a broadly responsive substrate such as 520 MMP FRET Substrate 14 is valuable for measuring cumulative MMP activity. 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 substrate, fluorescence is quenched due to close proximity between donor and quencher. Upon enzymatic cleavage: The peptide bond is hydrolyzed. The donor and quencher are separated. Fluorescence intensity increases proportionally to proteolytic activity. This design allows continuous real-time monitoring of MMP-mediated cleavage with high sensitivity and minimal background. Research Applications 1. Broad MMP Activity Profiling The substrate supports functional assessment of total MMP activity in: Cell culture supernatants Tissue homogenates Inflammatory exudates Serum or plasma samples 2. Extracellular Matrix Remodeling Studies It is well suited for investigating ECM degradation in models of fibrosis, wound healing, tumor invasion, and cartilage destruction. 3. Inflammatory Signaling Research MMP-3, -9, and -12 play roles in inflammatory cascades and cytokine processing. This substrate enables mechanistic analysis of proteolytic contributions to inflammation. 4. Inhibitor Screening and Drug Discovery The robust fluorescence readout is compatible with medium- to high-throughput screening platforms for evaluating MMP inhibitors. Reduced fluorescence indicates effective protease inhibition. Advantages Broad reactivity across major MMP subtypes High signal-to-background ratio due to FRET design Real-time kinetic measurement capability Suitable for complex biological samples Compatible with standard fluorescence microplate readers Experimental Considerations Assay conditions should include zinc and calcium ions required for MMP catalytic activity. Latent pro-MMPs may require activation (e.g., with APMA) before analysis. Use of selective MMP inhibitors can help confirm specificity when profiling mixed protease samples.