5-FAM MMP FRET Peptide Fluorescence Standard

Product Name: 5-FAM MMP FRET Peptide Fluorescence Standard

Sequence One Letter Code: 5-FAM-PL

Sequence Three Letter Code: 5-FAM-Pro-Leu-OH

Chemical Formula:C32H30N2O9

Molecular Weight: 586.6

Purity: 95%

Form: Lyophilized

Storage Conditions: - 20 °C Protected from light

Research Area: Peptide Series

Conjugation: Conjugated

Conjugation Type: Fluorescent dyes

Code Nacres: NA.26

Application: 5-FAM MMP FRET Peptide Fluorescence Standard is a fluorophore-labeled peptide designed for calibration and quantitative normalization in matrix metalloproteinase assays. Structurally analogous to the fluorescent fragment generated upon cleavage of 520 MMP FRET substrates, it contains the 5-FAM fluorophore without a quencher moiety. This peptide is used to generate fluorescence standard curves for accurate quantification of MMP activity. Fluorescence detection at excitation/emission 494/521 nm ensures direct compatibility with FRET-based MMP assay systems used in extracellular matrix and tissue remodeling research.

Current Research: 5-FAM MMP FRET Peptide Fluorescence Standard is a fluorophore-labeled reference peptide developed for calibration and quantitative normalization in matrix metalloproteinase (MMP) activity assays. The peptide is structurally analogous to the fluorescent fragment generated following proteolytic cleavage of 520 MMP FRET substrates, but contains the 5-carboxyfluorescein (5-FAM) fluorophore without an associated quencher moiety. Because it mimics the liberated fluorescent product of FRET-based MMP substrates, this standard enables accurate conversion of fluorescence intensity into molar concentration, supporting quantitative enzymatic analysis. Functional Rationale In intact FRET substrates, fluorescence is suppressed due to proximity between donor and quencher. Upon MMP-mediated cleavage: The quencher is separated from the fluorophore Free 5-FAM–containing fragments are released Fluorescence intensity increases proportionally to enzymatic activity The 5-FAM MMP Fluorescence Standard represents this cleaved fluorescent fragment, allowing generation of standard curves to correlate fluorescence units with absolute product concentration. Spectral Properties Excitation: ~494 nm Emission: ~521 nm These spectral characteristics ensure direct compatibility with commonly used fluorescence microplate readers and FRET-based MMP assay systems. Research Applications 1. Standard Curve Generation The peptide is used to prepare calibration curves for determining the concentration of cleaved fluorescent product in MMP assays. This enables conversion of raw fluorescence values into: Molar product formation Enzyme velocity (V₀) Specific activity 2. Assay Normalization It supports normalization across experimental runs, instruments, or sample sets, improving reproducibility and quantitative accuracy. 3. Instrument Calibration The standard can be used to verify instrument performance and detector sensitivity at the relevant wavelength pair. 4. Inhibitor Evaluation Studies In inhibitor screening workflows, accurate quantification of residual enzymatic activity requires reliable fluorescence calibration, which this standard provides. Advantages Direct structural correspondence to cleaved FRET substrate products Eliminates variability associated with incomplete cleavage assumptions Compatible with all 520 nm MMP FRET assay formats Improves quantitative precision and inter-assay comparability Experimental Considerations Standard curves should be prepared under identical buffer conditions as the assay to ensure accurate fluorescence correlation. Calibration should cover the anticipated range of product formation. Protection from prolonged light exposure is recommended to preserve fluorophore stability.