Ac-Phe-Arg-AMC HCl is a fluorogenic dipeptide used in monitoring protease activity through AMC release. The aromatic and basic residues enable precise recognition of enzymatic pockets. Researchers apply it in kinetic profiling, inhibitor screening, and specificity mapping. Its acetylated N-terminus and hydrochloride form support stable assay performance.
CAT No: R2257
CAS No:177028-04-5
Synonyms/Alias:Ac-Phe-Arg-AMC HCl;177028-04-5;(2S)-2-[[(2S)-2-acetamido-3-phenylpropanoyl]amino]-5-(diaminomethylideneamino)-N-(4-methyl-2-oxochromen-7-yl)pentanamide;hydrochloride;Ac-Phe-Arg-AMC . HCl;Ac-Phe-Arg-AMC hydrochloride;FA110473;
Ac-Phe-Arg-AMC HCl is a synthetic peptide substrate widely recognized in biochemical research for its utility in enzymatic activity assays, particularly those involving serine proteases such as trypsin-like enzymes. Structurally, it comprises an acetylated phenylalanine-arginine dipeptide linked to 7-amino-4-methylcoumarin (AMC), with the hydrochloride salt improving solubility and handling. Upon enzymatic cleavage at the arginine-AMC bond, the AMC moiety is released, yielding a highly fluorescent signal that can be quantitatively measured. This property makes the compound an essential tool for kinetic studies, inhibitor screening, and the detailed characterization of proteolytic mechanisms in vitro.
Enzyme Activity Assays: The primary application of Ac-Phe-Arg-AMC HCl is as a fluorogenic substrate in the quantitative measurement of serine protease activity, especially trypsin-like enzymes. Researchers utilize its specific cleavage site to monitor protease function in real time, as the release of AMC generates a robust fluorescent signal proportional to enzymatic turnover. This enables sensitive and high-throughput assessment of protease kinetics, facilitating the study of enzyme specificity, catalytic efficiency, and substrate preferences across diverse biological samples.
Protease Inhibitor Screening: The compound serves a vital role in the evaluation of protease inhibitors within drug discovery pipelines and mechanistic enzymology. By incorporating Ac-Phe-Arg-AMC HCl into assay platforms, scientists can rapidly assess the potency and selectivity of candidate inhibitors based on their ability to block substrate cleavage and subsequent AMC fluorescence. This approach supports the identification and optimization of novel inhibitory molecules, advancing research into protease regulation and potential therapeutic targets.
Biochemical Pathway Elucidation: In cellular and molecular biology, the substrate is instrumental for dissecting proteolytic pathways and mapping enzyme cascades. Its selective recognition by trypsin-like proteases allows for the tracking of proteolytic events within complex mixtures, such as cell lysates or tissue extracts. By quantifying changes in fluorescence, investigators can infer the presence, activity levels, and regulatory dynamics of key proteases, contributing to a deeper understanding of biological processes such as cell signaling, apoptosis, and protein turnover.
Assay Development and Optimization: The robust and reproducible fluorescent readout generated by Ac-Phe-Arg-AMC HCl makes it an invaluable standard in the development and validation of high-throughput screening assays. Analytical laboratories and assay developers employ the substrate to calibrate detection systems, optimize assay conditions, and establish performance benchmarks for automated platforms. Its well-characterized cleavage kinetics and compatibility with microplate readers enhance assay reliability and facilitate the translation of research protocols to industrial or diagnostic contexts.
Educational and Methodological Research: Beyond applied research, the compound is frequently adopted in educational and methodological studies to illustrate fundamental concepts in enzymology and biochemistry. Its straightforward mechanism of action, combined with easily detectable fluorescence, allows instructors and trainees to demonstrate principles such as enzyme-substrate interactions, Michaelis-Menten kinetics, and the effects of inhibitors in a laboratory setting. This supports hands-on learning and the dissemination of best practices in biochemical assay design and data interpretation.
2. TMEM16F and dynamins control expansive plasma membrane reservoirs
5. Autoinhibition and phosphorylation-induced activation of phospholipase C-γ isozymes
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