Recombinant Mouse Aminopeptidase N/CD13 Protein, CF

R&D Systems | Catalog # 2335-ZN

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Aminopeptidase N/CD13 Protein (2335-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

P97449

Applications

Enzyme Activity
View all Aminopeptidase N/CD13 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Aminopeptidase N/CD13 protein
Lys69-Ser966, with a C-terminal 10-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

Lys69

Predicted Molecular Mass

104 kDa

SDS-PAGE

111-123 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Ala-7-amido-4-methylcoumarin (Ala-AMC).
The specific activity is >1,000 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2335-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 N/CD13

The mouse ANPEP gene encodes Aminopeptidase N (APN), which is also known as microsomal aminopeptiase, alanyl aminopeptidase, Aminopeptidase M, CD13, or membrane protein p161 (1-3). The deduced amino acid sequence of mouse APN consists of a short cytoplasmic tail (residues 2 to 8), a transmembrane region (residue 9 to 32), a Ser/Thr rich region and a zinc metalloprotease domain (residues 69 to 966). Widely expressed in many cells, tissues and species, APN cleaves the N-terminal amino acids from bioactive peptides, leading to their inactivation or degradation. The roles of APN in many fields, such as neuroscience, hematopoeitic cells, immune system, angiogenesis, cancer and viral infection, have been reviewed (4).

References

  1. Chen, H. et al. (1996) J. Immunol. 157:2593.
  2. Larsen, S.L. et al. (1996) J. Exp. Med. 184:183.
  3. Hansen, A.S. et al. (1993) Eur. J. Immunol. 23:2358.
  4. Turner, A.J. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 289 Academic Press, San Diego.

Alternate Names

Aminopeptidase M, ANPEP, APN, CD13, gp150, PEPN

Entrez Gene IDs

290 (Human); 16790 (Mouse); 397520 (Porcine); 102115663 (Cynomolgus Monkey)

Gene Symbol

ANPEP

UniProt

P97449

Additional Aminopeptidase N/CD13 Products

Product Documents for Recombinant Mouse Aminopeptidase N/CD13 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 Mouse Aminopeptidase N/CD13 Protein, CF

For research use only

Citations for Recombinant Mouse Aminopeptidase N/CD13 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Aminopeptidase N/CD13 Protein, CF (2335-ZN):

Materials
  • Assay Buffer: 50 mM Tris, pH 7.0
  • Recombinant Mouse Aminopeptidase N/CD13 (rmCD13) (Catalog # 2335-ZN)
  • Substrate: H-Ala-AMC (Bachem, Catalog # I-1410), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmCD13 to 1 µg/mL in Assay Buffer.
  2. Incubate at 37 °C for one hour (fully activates enzyme).
  3. Dilute rmCD13 to 0.2 ng/µL in Assay Buffer.
  4. Dilute Substrate to 200 µM in Assay Buffer.
  5. Load 50 µL of 0.2 ng/µL rmCD13 in a plate, 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.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. 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-Courmarin (AMC) (Sigma, Catalog # A-9891).

Per Well:
  • rmCD13 0.010 µg
  • Substrate: 100 µM

FAQs

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