L-Ala-Ala-Phe-7-Amido-4-methylcoumarin Tfa salt links a tripeptide to a fluorogenic reporter, enabling quantitative studies of protease activity. Phenylalanine enhances hydrophobic recognition, shaping enzymatic interactions. Researchers analyze its fluorescence output during cleavage reactions. Uses include kinetic profiling, substrate evaluation, and assay development.
CAT No: R2475
CAS No:62037-42-7
Synonyms/Alias:Ala-Ala-Phe-AMC (TFA);H-AAF-AMC,TFA;62037-42-7;L-Ala-Ala-Phe-7-Amido-4-methylcoumarin Tfa salt;HY-137164A;CS-1050127;(S)-2-((S)-2-((S)-2-aminopropanamido)propanamido)-N-(4-methyl-2-oxo-2H-chromen-7-yl)-3-phenylpropanamide 2,2,2-trifluoroacetate;
L-Ala-Ala-Phe-7-Amido-4-methylcoumarin Tfa salt is a synthetic peptide substrate widely utilized in biochemical and molecular biology research for its unique fluorogenic properties. Composed of a tripeptide sequence—L-alanine-alanine-phenylalanine—conjugated to the 7-amido-4-methylcoumarin (AMC) fluorophore, this compound enables sensitive detection of protease activity. The Tfa (trifluoroacetate) salt form enhances its solubility and stability, making it an ideal candidate for enzymatic assays and kinetic studies. Upon enzymatic cleavage, the AMC moiety is released, resulting in a measurable fluorescent signal, which allows for real-time monitoring of proteolytic reactions. Its application spans across various research domains, providing a reliable tool for investigating enzyme specificity, inhibitor screening, and substrate profiling. Researchers value the compound for its consistent performance, high sensitivity, and compatibility with a range of assay platforms, including microplate readers and fluorescence spectrophotometers.
Enzyme Activity Assays: L-Ala-Ala-Phe-7-Amido-4-methylcoumarin Tfa salt is extensively used as a substrate in fluorometric assays to quantify the activity of proteases, particularly those with chymotrypsin-like specificity. In these assays, the peptide bond adjacent to the AMC group is selectively cleaved by target enzymes, resulting in a rapid increase in fluorescence intensity. This enables researchers to monitor enzyme kinetics in real time and calculate important parameters such as Km and Vmax. The substrate's high sensitivity allows for the detection of even low levels of proteolytic activity, making it indispensable for characterizing enzyme function in various biological samples.
Protease Inhibitor Screening: The AMC-conjugated peptide serves as a powerful tool in high-throughput screening platforms for the identification and evaluation of protease inhibitors. By incorporating the substrate into automated assay systems, researchers can rapidly assess the inhibitory potential of small molecules, peptides, or natural products. The fluorescence-based readout provides a robust, quantitative measure of enzyme inhibition, facilitating the discovery of novel compounds for research and therapeutic development. Its compatibility with microplate formats and automated liquid handling systems streamlines the screening process, enabling efficient analysis of large compound libraries.
Substrate Specificity Profiling: L-Ala-Ala-Phe-AMC is frequently employed to investigate the substrate preferences of various proteolytic enzymes. By comparing the rate of AMC release across different peptide sequences, researchers can delineate the structural determinants of enzyme-substrate recognition. This information is crucial for understanding the molecular mechanisms underlying protease function and for designing more selective substrates or inhibitors. The fluorogenic nature of the compound allows for rapid, high-throughput analysis, supporting comprehensive profiling studies in both academic and industrial research settings.
Cellular and Tissue Extract Analysis: The peptide substrate is also utilized in the analysis of protease activity within complex biological samples, such as cell lysates or tissue extracts. By adding the substrate to these samples, researchers can assess endogenous protease activities under physiological or experimental conditions. The resulting fluorescence provides a sensitive and quantitative measure of enzyme function, enabling the study of proteolytic processes involved in cellular regulation, signal transduction, and pathological states. Its use in multiplexed assays further allows for simultaneous monitoring of multiple enzymatic activities within a single experiment.
Biochemical Mechanism Studies: Researchers leverage L-Ala-Ala-Phe-7-Amido-4-methylcoumarin Tfa salt to elucidate the catalytic mechanisms of serine and cysteine proteases. By analyzing the kinetics of substrate hydrolysis, it is possible to infer details about enzyme active sites, transition states, and catalytic residues. This mechanistic insight informs the rational design of selective inhibitors and enhances our understanding of enzyme regulation in biological systems. The compound's well-defined structure and predictable cleavage pattern make it a preferred choice for mechanistic studies that require precise measurement of proteolytic events.
In summary, L-Ala-Ala-Phe-7-Amido-4-methylcoumarin Tfa salt stands out as a versatile and highly effective fluorogenic substrate in the field of protease research. Its applications encompass enzyme activity assays, inhibitor screening, substrate specificity profiling, analysis of cellular extracts, and mechanistic investigations. By enabling sensitive and quantitative detection of proteolytic activity, the compound supports a wide range of scientific inquiries, from basic enzymology to advanced drug discovery efforts. Its reliability, ease of use, and adaptability to various experimental formats have established it as an essential tool for researchers seeking to unravel the complexities of protease function and regulation.
2. Cationic cell-penetrating peptides are potent furin inhibitors
4. SERS spectrum of the peptide thymosin‐β4 obtained with Ag nanorod substrate
If you have any peptide synthesis requirement in mind, please do not hesitate to contact us at . We will endeavor to provide highly satisfying products and services.
Creative Peptides is a trusted CDMO partner specializing in high-quality peptide synthesis, conjugation, and manufacturing under strict cGMP compliance. With advanced technology platforms and a team of experienced scientists, we deliver tailored peptide solutions to support drug discovery, clinical development, and cosmetic innovation worldwide.
From custom peptide synthesis to complex peptide-drug conjugates, we provide flexible, end-to-end services designed to accelerate timelines and ensure regulatory excellence. Our commitment to quality, reliability, and innovation has made us a preferred partner across the pharmaceutical, biotechnology, and personal care industries.
Read More