Recombinant Human Aminopeptidase A/ENPEP Protein, CF

R&D Systems | Catalog # 2499-ZN

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Aminopeptidase A/ENPEP Protein (2499-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

AAA35522

Applications

Enzyme Activity
View all Aminopeptidase A/ENPEP Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Aminopeptidase A/ENPEP protein
Arg41-Gly957, with an N-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His

Predicted Molecular Mass

106 kDa

SDS-PAGE

145 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Glu-7-amido-4-methylcoumarin (Glu-AMC).
The specific activity is > 2,000 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2499-ZN
Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Aminopeptidase A/ENPEP

The human ENPEP gene encodes aminopeptidase A (APA), which is also known as glutamyl aminopeptidase (EAP), aspartate aminopeptidase, angiotensinase A, Ca2+-activated glutamate aminopeptidase, membrane aminopeptidase II and the BP-1/6C3 antigen (1). The various names reflect its substrate specificity (with a preference for Glu, Asp and angiotensin II as one of the physiological substrates), Ca2+-dependence, cellular location (type II membrane protein) and immunological reactivity to the antibody BP-1/6C3. The deduced amino acid sequence of human ENPEP consists of a short cytoplasmic tail (residues 1 to 17), a transmembrane region (residues 18 to 40), and a long ectodomain (residues 41 to 957) (2, 3). In addition to the N-terminal zinc metalloprotease domain, the ectodomain also contains the C-terminal region, which functions as an intramolecular chaperone required for the correct folding, cell surface expression and activity (4). The purified recombinant human ENPEP consists of the entire ectodomain and is active in the assay as described in Activity Assay Protocol.

References

  1. Wang, J. and M.D. Cooper (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) p. 299, Academic Press, San Diego.
  2. Nanus, D.M. et al. (1993) Proc. Natl. Acad. Sci. USA 90:7069.
  3. Li, L. et al. (1993) Genomics 17:657.
  4. Rozenfeld, R. et al. (2004) J. Biol. Chem. 279:43285.

Alternate Names

CD249, ENPEP

Entrez Gene IDs

2028 (Human); 13809 (Mouse)

Gene Symbol

ENPEP

UniProt

AAA35522

Additional Aminopeptidase A/ENPEP Products

Product Documents for Recombinant Human Aminopeptidase A/ENPEP Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Aminopeptidase A/ENPEP Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Human Aminopeptidase A/ENPEP Protein, CF

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Protocols

View specific protocols for Recombinant Human Aminopeptidase A/ENPEP Protein, CF (2499-ZN):

Materials
  • Assay Buffer: 25 mM Tris, 50 mM CaCl2, 0.2 M NaCl, pH 8.0
  • Recombinant Human Aminopeptidase A/ENPEP (rhENPEP) (Catalog # 2499-ZN)
  • Substrate: Glu-AMC (Bachem, Catalog # I-1180), 10 mM stock in 30 mM HCl
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhENPEP to 0.4 ng/µL in Assay Buffer.
  2. Dilute Substrate Glu-AMC to 200 µM in Assay Buffer.
  3. Load into plate 50 µL of 0.4 ng/µL rhENPEP, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, 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 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:
  • rhENPEP: 0.020 μg
  • Substrate: 100 µM

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