Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate

Substrate for ECE-1, ACE, Cathepsin A, Cathepsin X/Z/P, Neprilysin, and Insulysin
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R&D Systems Fluorogenic Peptide Substrates
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Citations (17)
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Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate Summary

Product Specifications

>95%, by HPLC.
The peptide substrate contains a highly fluorescent 7-methoxycoumarin group that is efficiently quenched by resonance energy transfer to the 2,4-dinitrophenyl group. It can be used to measure the activities of peptidases that are capable of cleaving an amide bond between the fluorescent group and the quencher group, causing an increase in fluorescence. It is an excellent substrate for endothelin-converting enzyme-1 (ECE-1) and neprilysin. The cleavage site by ECE-1 is the peptide bond between Ala and Phe (Johnson, G.D. and Ahn, K., 2000, Anal. Biochem. 286:112). The substrate is derived from bradykinin. It is also an excellent substrate for ACE and active Cathepsin A and X/Z/P.

Mca: (7-Methoxycoumarin-4-yl)acetyl, Dnp: 2, 4-Dinitrophenyl.
Predicted Molecular Mass
1388 Da

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Stock solution at 4 mM or 6.26 mg/mL (0.16 mL) dimethyl sulfoxide (DMSO).
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Samples are stable for up to twelve months from date of receipt at -20° C to -70° C. The substrate can be aliquoted and stored -20° C to -70° C in a manual defrost freezer for six months. Avoid repeated freeze-thaw cycles. Protect from exposure to direct light
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Citations for Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

17 Citations: Showing 1 - 10
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  1. Medium-Chain Length Fatty Acids Enhance Abeta Degradation by Affecting Insulin-Degrading Enzyme
    Authors: J Mett, AA Lauer, D Janitschke, LV Griebsch, EL Theiss, HS Grimm, H Koivisto, H Tanila, T Hartmann, MOW Grimm
    Cells, 2021-10-29;10(11):.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay
  2. High affinity binding of SARS-CoV-2 spike protein enhances ACE2 carboxypeptidase activity.
    Authors: Lu J, Sun P
    J Biol Chem, 2020-10-29;295(52):18579-18588.
    Species: Human
    Sample Types: Peptide
    Applications: Bioassay
  3. Cathepsin A contributes to left ventricular remodeling by degrading extracellular superoxide dismutase in mice
    Authors: M Hohl, M Mayr, L Lang, AG Nickel, J Barallobre, X Yin, T Speer, SR Selejan, C Goettsch, K Erb, C Fecher-Tro, JC Reil, B Linz, S Ruf, T Hübschle, C Maack, M Böhm, T Sadowski, D Linz
    J. Biol. Chem., 2020-07-09;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
    Applications: Bioassay
  4. High affinity binding of SARS-CoV-2 spike protein enhances ACE2 carboxypeptidase activity
    Authors: J Lu, PD Sun
    bioRxiv, 2020-07-01;0(0):.
    Applications: Enzyme Assay
  5. Female Mice Exposed to Postnatal Neglect Display Angiotensin II-Dependent Obesity-Induced Hypertension
    Authors: C Dalmasso, JR Leachman, CM Ensor, FB Yiannikour, JF Giani, LA Cassis, AS Loria
    J Am Heart Assoc, 2019-11-22;8(23):e012309.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  6. Neuroprotection induced by Navbeta2?knockdown in APP/PS1 transgenic neurons is associated with NEP regulation
    Authors: T Hu, SS Li, MN Lu, L Zhang, B Chen, R Mao, R Mei, YX Tan, S Li, YB Xiyang
    Mol Med Rep, 2019-06-20;20(2):2002-2011.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Bioassay
  7. The selectivity of galardin and an azasugar-based hydroxamate compound for human matrix metalloproteases and bacterial metalloproteases
    Authors: I Sylte, R Dawadi, N Malla, S von Hofste, TM Nguyen, AI Solli, E Berg, OA Adekoya, G Svineng, JO Winberg
    PLoS ONE, 2018-08-03;13(8):e0200237.
    Species: Bacteria
    Sample Types: Protein
  8. Functional requirement for human pitrilysin metallopeptidase 1 arginine 183, mutated in amyloidogenic neuropathy
    Authors: JE Smith-Carp, BJ Alper
    Protein Sci., 2018-02-23;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  9. The anti-oxidant xanthorrhizol prevents amyloid-?-induced oxidative modification and inactivation of neprilysin
    Authors: CS Lim, JS Han
    Biosci. Rep., 2018-02-02;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  10. Vitamin D and Its Analogues Decrease Amyloid-? (A?) Formation and Increase A?-Degradation
    Authors: MOW Grimm, A Thiel, AA Lauer, J Winkler, J Lehmann, L Regner, C Nelke, D Janitschke, C Benoist, O Streidenbe, H Stötzel, K Endres, C Herr, C Beisswenge, HS Grimm, R Bals, F Lammert, T Hartmann
    Int J Mol Sci, 2017-12-19;18(12):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Activity Assay
  11. Characterization of insulin degrading enzyme and other amyloid-beta degrading proteases in human serum: a role in Alzheimer's disease?
    Authors: Liu Z, Zhu H, Fang G, Walsh K, Mwamburi M, Wolozin B, Abdul-Hay S, Ikezu T, Leissring M, Qiu W
    Oncogene, 2012-01-01;29(2):329-40.
    Species: Human
    Sample Types: Serum
    Applications: Enzyme Assay Substrate
  12. HIV-1 reduces Abeta-degrading enzymatic activities in primary human mononuclear phagocytes.
    Authors: Lan X, Xu J, Kiyota T, Peng H, Zheng JC, Ikezu T
    J. Immunol., 2011-05-06;186(12):6925-32.
    Species: Human
    Sample Types: Cell Lysates
  13. Expression and functional profiling of neprilysin, insulin-degrading enzyme, and endothelin-converting enzyme in prospectively studied elderly and Alzheimer's brain.
    Authors: Wang S, Wang R, Chen L, Bennett DA, Dickson DW, Wang DS
    J. Neurochem., 2010-07-30;115(1):47-57.
    Species: Human
    Sample Types: Tissue Homogenates
  14. Intra- versus extracellular effects of microglia-derived cysteine proteases in a conditioned medium transfer model.
    Authors: Wendt W, Schulten R, Stichel CC, Lubbert H
    J. Neurochem., 2009-07-17;110(6):1931-41.
    Species: Mouse
    Sample Types: Cell Lysates
  15. ACE mediates ventilator-induced lung injury in rats via angiotensin II but not bradykinin.
    Authors: Wosten-van Asperen RM, Lutter R, Haitsma JJ, Merkus MP, van Woensel JB, van der Loos CM, Florquin S, Lachmann B, Bos AP
    Eur. Respir. J., 2007-10-24;31(2):363-71.
    Species: Rat
    Sample Types: BALF
  16. Immunocapture-based fluorometric assay for the measurement of neprilysin-specific enzyme activity in brain tissue homogenates and cerebrospinal fluid.
    Authors: Miners JS, Verbeek MM, Rikkert MO, Kehoe PG, Love S
    J. Neurosci. Methods, 2007-08-25;167(2):229-36.
    Species: Human
    Sample Types: Tissue Homogenates
  17. Human Opiorphin, a natural antinociceptive modulator of opioid-dependent pathways.
    Authors: Wisner A, Dufour E, Messaoudi M, Nejdi A, Marcel A, Ungeheuer MN, Rougeot C
    Proc. Natl. Acad. Sci. U.S.A., 2006-11-13;103(47):17979-84.
    Species: Human
    Sample Types: Recombinant Protein


  1. Where does Cathepsin A cleave Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate, Catalog # ES005?

    • Although the QC assay conditions provided on our recombinant Cathepsin A datasheets should favor carboxypeptidase activity, it is possible that there is more than one site in the ES005 peptide recognized and cleaved by Cathepsin A.  We have not performed verification experiments to confirm the cleavage site preferred in our reaction conditions.

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Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate
By Shrinidh Joshi on 08/04/2016

ACE activity was determined following incubation with intramolecularly quenched synthetic ACE specific substrate Mca-RPGFSAFK (Dnp)-OH (R&D systems). In the case of cell lysates, 10 µg of total protein was assayed for activity in a buffer with the following composition: 50 mM MES (4-morpholineethanesulphonic acid), 300 mM NaCl, 10 µm ZnCl2 and 0.01% Triton X-100, pH 6.5. Reaction was initiated by the addition of 5×10−5 M substrate. Where applicable, recombinant enzymes were used at a concentration of 0.01 µg per reaction. The fluorescence measurements were performed in the black microtiter plates (Costar) in a total volume of 100 µl. The plates were read using a fluorescence plate reader SpectraMax M5 (Molecular Devices) at an excitation wavelength 320 nm and emission wavelength 405 nm Fluorescence resulting from the substrate hydrolysis increased with time, and achieved maximum by one h with recombinant enzyme however with cell/tissue lysates the maximum fluorescence was observed by four hours of incubation. Therefore fluorescence recorded at one and four hours of reaction time was taken for calculation of percent enzyme inhibition, when using recombinant enzymes and cell/tissue lysates, respectively. ACE activity was defined as the ACE inhibitor, captopril-sensitive fluorescence, and were expressed as percent inhibition.