Mca-DEVDAPK(Dnp)-OH | CAS 189696-20-6

Catalog Number	A18-0042
Category	Fluorescent Enzyme Substrates
Molecular Formula	C50H62N10O22
Molecular Weight	1155.1

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Product Introduction

Mca-DEVDAPK(Dnp)-OH is a synthetic fluorogenic peptide probe that features a 7-methoxycoumarin (Mca) fluorophore and a dinitrophenyl (Dnp) quencher. This compound is designed to participate in fluorescence resonance energy transfer (FRET) assays, where cleavage by specific proteases releases the fluorophore, resulting in a measurable increase in fluorescence. It is commonly used in biochemical research to study enzyme kinetics and caspase activity, providing insights into apoptotic pathways and proteolytic processes.

Chemical Information
Product Specification
Application

Chemical Information

Synonyms	Caspase-3 Fluorogenic Substrate III; CPP32 Fluorogenic Substrate III; 7-Methoxycoumarin-4-acetyl-DEVDAPK(Dnp)-OH; N-[(7-methoxy-2-oxo-2H-1-benzopyran-4-yl)acetyl]-L-α-aspartyl-L-α-glutamyl-L-valyl-L-α-aspartyl-L-alanyl-L-prolyl-N6-(2,4-dinitrophenyl)-L-lysine
Purity	≥95%
SMILES	CC(C)C(C(=O)NC(CC(=O)O)C(=O)NC(C)C(=O)N1CCCC1C(=O)NC(CCCCNC2=C(C=C(C=C2)[N+](=O)[O-])[N+](=O)[O-])C(=O)O)NC(=O)C(CCC(=O)O)NC(=O)C(CC(=O)O)NC(=O)CC3=CC(=O)OC4=C3C=CC(=C4)OC
InChI	InChI=1S/C50H62N10O22/c1-24(2)43(57-44(69)31(14-15-39(62)63)54-46(71)33(22-40(64)65)53-38(61)18-26-19-42(68)82-37-21-28(81-4)11-12-29(26)37)48(73)56-34(23-41(66)67)45(70)52-25(3)49(74)58-17-7-9-35(58)47(72)55-32(50(75)76)8-5-6-16-51-30-13-10-27(59(77)78)20-36(30)60(79)80/h10-13,19-21,24-25,31-35,43,51H,5-9,14-18,22-23H2,1-4H3,(H,52,70)(H,53,61)(H,54,71)(H,55,72)(H,56,73)(H,57,69)(H,62,63)(H,64,65)(H,66,67)(H,75,76)/t25-,31-,32-,33-,34-,35-,43-/m0/s1
InChIKey	YOUVSUDOGAVRQX-AEAFXXELSA-N

Product Specification

Storage

Store at -20°C

Application

Mca-DEVDAPK(Dnp)-OH is a quenched fluorogenic peptide substrate designed for monitoring protease activity via fluorescence readout. The molecule incorporates a DEVD recognition sequence commonly used in caspase assays and a Dnp (dinitrophenyl) quencher motif that suppresses fluorescence until proteolytic processing separates the fluorophore and quencher elements. This format is widely used in chemical biology and assay development workflows where enzyme-dependent signal generation is required.

1. Caspase Activity Assays

Mca-DEVDAPK(Dnp)-OH is used in fluorescence-based caspase activity measurements, particularly for DEVD-directed protease activity in cell lysates and purified enzyme preparations. Researchers employ the substrate in microplate formats to quantify time-dependent cleavage by tracking fluorescence increase after proteolysis relieves quenched emission. The peptide design supports straightforward assay workflows for screening protease modulators, optimizing buffer conditions, and comparing relative activity across experimental conditions.

2. Enzyme Kinetics Screening

Mca-DEVDAPK(Dnp)-OH supports kinetic studies where reaction rates are extracted from fluorescence versus time traces. In protease-focused assay development, the quenched peptide format helps reduce background from free fluorophore signal, enabling more reliable monitoring of cleavage progression during optimization of enzyme concentration, incubation time, and assay throughput. This reagent is commonly handled by assay development teams in academic core facilities and industrial R&D groups building enzymatic readouts for mechanistic studies and inhibitor profiling.

3. Protease-Dependent Signal Generation

Mca-DEVDAPK(Dnp)-OH is applied as a fluorescence turn-on substrate to generate enzyme-dependent signal in biochemical studies of apoptosis-related protease pathways. Investigators use the DEVD-directed cleavage behavior to link protease activity to experimental perturbations such as genetic knockdowns, lysate preparation changes, or chemical treatments in controlled laboratory settings. The quenched architecture makes it useful for workflows where minimizing baseline fluorescence is important for interpreting cleavage-dependent changes across samples.

4. Cell Lysate Fluorogenic Readouts

Mca-DEVDAPK(Dnp)-OH is frequently used with cell lysates to assess protease activity in complex biological mixtures while maintaining a fluorescence-based readout. Researchers can run parallel reactions under defined lysis and incubation conditions to compare relative protease activity between samples, including fractionated lysates and standardized protein inputs. The peptide substrate format aligns with routine fluorescence plate-reader measurements used in molecular biology laboratories for pathway activity monitoring and assay condition validation.