Ac-DEVD-AFC is a fluorogenic substrate (λex=400 nm, λem=530 nm).
CAT No: R1146
CAS No:201608-14-2
Synonyms/Alias:Ac-DEVD-AFC;Ac-Asp-Glu-Val-Asp-AFC;201608-14-2;MFCD01310970;N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-trifluoromethylcoumarin;(4S)-4-[[(2S)-2-acetamido-3-carboxypropanoyl]amino]-5-[[(2S)-1-[[(2S)-3-carboxy-1-oxo-1-[[2-oxo-4-(trifluoromethyl)chromen-7-yl]amino]propan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-5-oxopentanoic acid;Ac-Asp-Glu-Val-Asp-7-amino-4-trifluoromethylcoumarin;SCHEMBL1180114;DTXSID80942217;GZDRODOYEFEHGG-NUDCOPPTSA-N;Caspase-3 Substrate (Fluorogenic);HY-P1005;CS-7683;BP-23699;DA-70473;MS-31264;F85555;N-Acetyl-Asp-Glu-Val-Asp-7-amino-4-(trifluoromethyl)coumarin;Ac-Asp-Glu-Val-Asp-AFC (AFC = 7-Amino-4-trifluoromethylcoumarin);N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-trifluoromethylcoumarin, >=90% (HPLC), powder;7-(2-Carboxyethyl)-4-(carboxymethyl)-2,5,8,11-tetrahydroxy-13-(hydroxy{[2-oxo-4-(trifluoromethyl)-2H-1-benzopyran-7-yl]imino}methyl)-10-(propan-2-yl)-3,6,9,12-tetraazapentadeca-2,5,8,11-tetraen-15-oic acid;L-a-Asparagine,N-acetyl-L-a-aspartyl-L-a-glutamyl-L-valyl-N-[2-oxo-4-(trifluoromethyl)-2H-1-benzopyran-7-yl]-;L-alpha-Asparagine, N-acetyl-L-alpha-aspartyl-L-alpha-glutamyl-L-valyl-N-[2-oxo-4-(trifluoromethyl)-2H-1-benzopyran-7-yl]- (9CI);
Ac-DEVD-AFC is a synthetic peptide substrate widely recognized for its utility in biochemical assays targeting caspase-3 and related proteases. Structurally, it consists of an acetylated tetrapeptide sequence (Asp-Glu-Val-Asp) conjugated to the fluorogenic moiety 7-amino-4-trifluoromethylcoumarin (AFC). Upon enzymatic cleavage at the DEVD site, the AFC group is released, producing a measurable fluorescent signal. This property makes Ac-DEVD-AFC an essential reagent for real-time monitoring of protease activity in diverse research settings, particularly those focused on apoptosis and cell death pathways.
Enzyme activity assays: One of the primary applications for Ac-DEVD-AFC is in kinetic and endpoint assays designed to quantify caspase-3 and related caspase activities. Researchers employ it to monitor enzymatic cleavage in cell lysates, purified enzyme preparations, or in vitro reconstituted systems. The release of the AFC fluorophore upon substrate cleavage enables sensitive and quantitative detection of proteolytic activity, facilitating the study of enzyme kinetics, substrate specificity, and inhibitor screening.
Apoptosis research: The DEVD sequence within this substrate is preferentially recognized by caspase-3, a critical executioner protease in programmed cell death. Utilization of Ac-DEVD-AFC in cellular or cell-free systems allows for the assessment of apoptotic progression by measuring caspase activation. Its application extends to evaluating the efficacy of apoptosis-inducing agents, dissecting signaling pathways, and validating mechanistic hypotheses regarding cell death regulation.
High-throughput screening: Owing to its robust fluorescence signal and specificity, Ac-DEVD-AFC is routinely incorporated into automated high-throughput screening platforms. This enables rapid evaluation of large compound libraries for potential modulators of caspase activity. The substrate's compatibility with microplate formats and standard fluorescence readers streamlines assay development, supporting drug discovery initiatives and functional genomics studies.
Mechanistic enzymology: The substrate's defined peptide sequence and fluorogenic properties make it valuable for mechanistic investigations into protease-substrate interactions. Researchers use it to probe active site preferences, determine catalytic efficiencies, and elucidate structural determinants of substrate recognition. Such studies contribute to a deeper understanding of protease function and inform the design of novel inhibitors or engineered enzymes.
Cell-based assay development: Ac-DEVD-AFC is frequently employed in the optimization and validation of cell-based assays for monitoring intracellular caspase activation. By enabling real-time and quantitative assessment of proteolytic activity within living cells, it supports the development of robust models for studying cellular stress responses, toxicology, and signal transduction. Its use in these contexts provides a powerful tool for linking biochemical events to functional outcomes in complex biological systems.
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