Recombinant Human ACE Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human ACE Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005). The specific activity is >1,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human ACE/CD143 protein
Leu30-Leu1261, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Leu30
Structure / Form
Recombinant Human ACE/CD143 Somatic Form is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Protein/Peptide Type
Recombinant Enzymes
Gene
ACE
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
143 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
160-180 kDa, reducing conditions
Publications
Read Publications using
929-ZN in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, ZnCl2 and Glycerol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM MES, pH 6.5
  • Recombinant Human ACE/CD143 Somatic Form (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
  1. Dilute rhACE to 0.4 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. 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.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  5. 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ACE Protein, CF

  • ACE
  • ACE1angiotensin converting enzyme, somatic isoform
  • angiotensin I converting enzyme (peptidyl-dipeptidase A) 1
  • carboxycathepsin
  • CD143 antigen
  • CD143
  • DCP
  • DCP1
  • DCP1angiotensin-converting enzyme
  • dipeptidyl carboxypeptidase 1
  • Dipeptidyl carboxypeptidase I
  • EC 3.2.1.-
  • EC 3.4.15.1
  • Kininase II
  • MGC26566
  • MVCD3
  • peptidase P
  • testicular ECA

Background

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).

  1. Soubrier, et al. (1988) Proc. Natl. Acad. Sci. USA 85:9386.
  2. Corvol, P. and T.A. Williams (1998) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. (eds): San Diego, Academic Press, p. 1066.
  3. Hu, et al. (2001) J. Biol. Chem. 276:47863.
  4. Kessler, et al. (2000) J. Biol. Chem. 275:26259.
  5. Eyries, et al. (2001) J. Biol. Chem. 276:5525.
  6. Alfalah, et al. (2001) J. Biol. Chem. 276:21105.

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Publications for ACE/CD143 (929-ZN)(6)

We have publications tested in 2 confirmed species: Human, Primate - Chlorocebus pygerythrus (Vervet Monkey).

We have publications tested in 2 applications: Bioassay, Enzyme Assay.


Filter By Application
Bioassay
(4)
Enzyme Assay
(2)
All Applications
Filter By Species
Human
(5)
Primate - Chlorocebus pygerythrus (Vervet Monkey)
(1)
All Species
Showing Publications 1 - 6 of 6.
Publications using 929-ZN Applications Species
NS Al Mulhim, PG Kehoe, JS Miners Divergence in the activity of the N- and C- catalytic domains of ACE1 - implications for the role of the renin-angiotensin system in Alzheimer&#039;s disease Acta Neuropathol Commun, 2019;7(1):57. 2019 [PMID: 31018864] (Bioassay, Human) Bioassay Human
AM South, PA Nixon, MC Chappell, DI Diz, GB Russell, HA Shaltout, TM O'Shea, LK Washburn Obesity is Associated with Higher Blood Pressure and Higher Levels of Angiotensin II but Lower Angiotensin-(1-7) in Adolescents Born Preterm J. Pediatr., 2018;0(0):. 2018 [PMID: 30404738] (Bioassay, Human) Bioassay Human
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 New perspectives in the renin-angiotensin-aldosterone system (RAAS) II: albumin suppresses angiotensin converting enzyme (ACE) activity in human. PLoS ONE, 2014;9(4):e87844. 2014 [PMID: 24691203] (Bioassay, Human) Bioassay Human
Roosterman D, Kempkes C, Cottrell GS, Padilla BE, Bunnett NW, Turck CW, Steinhoff M Endothelin-converting enzyme-1 degrades internalized somatostatin-14. Endocrinology, 2008;149(5):2200-7. 2008 [PMID: 18276747] (Enzyme Assay, Human) Enzyme Assay Human
Miners JS, Verbeek MM, Rikkert MO, Kehoe PG, Love S Immunocapture-based fluorometric assay for the measurement of neprilysin-specific enzyme activity in brain tissue homogenates and cerebrospinal fluid. J. Neurosci. Methods, 2008;167(2):229-36. 2008 [PMID: 17904641] (Enzyme Assay, Human) Enzyme Assay Human
de Lang A, Osterhaus AD, Haagmans BL Interferon-gamma and interleukin-4 downregulate expression of the SARS coronavirus receptor ACE2 in Vero E6 cells. Virology, 2006;353(2):474-81. 2006 [PMID: 16860835] (Bioassay, Primate - Chlorocebus pygerythrus (Vervet Monkey)) Bioassay Primate - Chlorocebus pygerythrus (Vervet Monkey)

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Bioinformatics

Gene Symbol ACE
Uniprot