Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate
R&D Systems | Catalog # ES005
Key Product Details
Conjugate
Applications
Product Specifications
Source
Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)-OH
Mca: (7-Methoxycoumarin-4-yl)acetyl, Dnp: 2, 4-Dinitrophenyl.
Purity
Predicted Molecular Mass
Activity
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.
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Formulation, Preparation, and Storage
ES005
| 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 |
Product Documents for Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate
Certificate of Analysis
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Product Specific Notices for Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate
For research use only
Citations for Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate
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Verified Customer | Posted 08/04/2016ACE 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.
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FAQs for Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate
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Q: Where does Cathepsin A cleave Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate, Catalog # ES005?
A: 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.