Recombinant Human ACE Protein, CF Summary
Product Specifications
Leu30-Leu1261, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
929-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, ZnCl2 and Glycerol. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 50 mM MES, pH 6.5
- Recombinant Human ACE (rhACE) (Catalog # 929-ZN)
- Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhACE to 0.4 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- In a plate load 50 µL of 0.4 ng/µL rhACE and start the reaction by adding 50 µL of 20 µM Substrate to the wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard MCA-P-L-OH (Bachem, Catalog # M-1975).
Per Well:- rhACE: 0.020 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: ACE/CD143
ACE (also known as peptidyl-dipetidase A) is a zinc metallopeptidase important for blood pressure control and water and salt metabolism (2). It cleaves the C-terminal dipeptide from angiotensin I to produce the potent vasopressor octapeptide angiotensin II and inactivates bradykinin by the sequential removal of two C-terminal dipeptides. In addition to the two physiological substrates, ACE cleaves C-terminal dipeptides from various oligopeptides with a free C-terminus. Because of its location and specificity, ACE plays additional roles in immunity, reproduction and neuropeptide regulation. For example, ACE degrades Alzheimer amyloid beta -peptide (A beta ), retards A beta aggregation, deposition, fibril formation, and inhibits cytotoxicity (3).
ACE is a type I membrane protein and exists in two isoforms (2). Somatic ACE, found in endothelial, epithelial and neuronal cells, comprises two highly similar domains called N- and C-domains, each of which contains the HExxH consensus sequence for zinc binding. Germinal ACE, found exclusively in the testes, comprises a single catalytically active domain identical to the C-domain of somatic ACE except for an N-terminal 67 residue germinal ACE-specific sequence. Physiological functions of the two tissue-specific isozymes are not interchangeable (4). For example, sperm-specific expression of the germinal ACE, not the somatic ACE, in ACE knockout male mice restored fertility.
Soluble ACE is present in many biological fluids, such as serum, seminal fluid, amniotic fluid and cerebrospinal fluid (2). The soluble ACE is derived from the membrane forms by actions of secretases or sheddases. The identities of the secretases have not been revealed, although they belong to the family of zinc metallopeptidases (5, 6).
- Soubrier, et al. (1988) Proc. Natl. Acad. Sci. USA 85:9386.
- Corvol, P. and T.A. Williams (1998) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. (eds): San Diego, Academic Press, p. 1066.
- Hu, et al. (2001) J. Biol. Chem. 276:47863.
- Kessler, et al. (2000) J. Biol. Chem. 275:26259.
- Eyries, et al. (2001) J. Biol. Chem. 276:5525.
- Alfalah, et al. (2001) J. Biol. Chem. 276:21105.
Citations for Recombinant Human ACE Protein, CF
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.
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Citations: Showing 1 - 7
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A Gamma-adapted subunit vaccine induces broadly neutralizing antibodies against SARS-CoV-2 variants and protects mice from infection
Authors: Coria, LM;Rodriguez, JM;Demaria, A;Bruno, LA;Medrano, MR;Castro, CP;Castro, EF;Del Priore, SA;Hernando Insua, AC;Kaufmann, IG;Saposnik, LM;Stone, WB;Prado, L;Notaro, US;Amweg, AN;Diaz, PU;Avaro, M;Ortega, H;Ceballos, A;Krum, V;Zurvarra, FM;Sidabra, JE;Drehe, I;Baqué, JA;Li Causi, M;De Nichilo, AV;Payes, CJ;Southard, T;Vega, JC;Auguste, AJ;Álvarez, DE;Flo, JM;Pasquevich, KA;Cassataro, J;
Nature communications
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA Standard -
Divergence in the activity of the N- and C- catalytic domains of ACE1 - implications for the role of the renin-angiotensin system in Alzheimer's disease
Authors: NS Al Mulhim, PG Kehoe, JS Miners
Acta Neuropathol Commun, 2019-04-24;7(1):57.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Obesity is Associated with Higher Blood Pressure and Higher Levels of Angiotensin II but Lower Angiotensin-(1-7) in Adolescents Born Preterm
Authors: AM South, PA Nixon, MC Chappell, DI Diz, GB Russell, HA Shaltout, TM O'Shea, LK Washburn
J. Pediatr., 2018-11-05;0(0):.
Species: Human
Sample Types: Urine
Applications: Bioassay -
New perspectives in the renin-angiotensin-aldosterone system (RAAS) II: albumin suppresses angiotensin converting enzyme (ACE) activity in human.
Authors: Fagyas M, Uri K, Siket I, Fulop G, Csato V, Darago A, Boczan J, Banyai E, Szentkiralyi I, Maros T, Szerafin T, Edes I, Papp Z, Toth A
PLoS ONE, 2014-04-01;9(4):e87844.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Endothelin-converting enzyme-1 degrades internalized somatostatin-14.
Authors: Roosterman D, Kempkes C, Cottrell GS, Padilla BE, Bunnett NW, Turck CW, Steinhoff M
Endocrinology, 2008-02-14;149(5):2200-7.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
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: Peptide
Applications: Enzyme Assay -
Interferon-gamma and interleukin-4 downregulate expression of the SARS coronavirus receptor ACE2 in Vero E6 cells.
Authors: de Lang A, Osterhaus AD, Haagmans BL
Virology, 2006-07-24;353(2):474-81.
Species: Primate - Chlorocebus pygerythrus (Vervet Monkey)
Sample Types: Whole Cells
Applications: Bioassay
FAQs
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Is ACE-1 the same as ACE?
Yes. ACE-1 is used as an alternative name, but not as commonly as ACE and CD143.
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