Recombinant Human Aminopeptidase N/CD13 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Aminopeptidase N/CD13 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Ala-7-amido-4-methylcoumarin (Ala-AMC). The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Aminopeptidase N/CD13 protein
Lys69-Lys967, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Lys69
Protein/Peptide Type
Recombinant Enzymes
Gene
ANPEP
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
104 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
131 kDa, reducing conditions
Publications
Read Publications using
3815-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 MES and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM Tris, pH 7.0
  • Recombinant Human Aminopeptidase N/CD13 (rhCD13) (Catalog # 3815-ZN)
  • Substrate: Ala-AMC (Bachem, Catalog # I-1410)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Spectramax Gemini EM by Molecular Devices) or equivalent

  1. Dilute rhCD13 to 0.2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load 50 µL of 0.2 µg/mL rhCD13 into 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 Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 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 7-Amino-4-Methyl Coumarin (Sigma, Catalog # A9891).

Per Well:
  • rhCD13: 0.010 µg
  • Substrate: 100 µM

Notes

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

Alternate Names for Recombinant Human Aminopeptidase N/CD13 Protein, CF

  • alanyl (membrane) aminopeptidase
  • Alanyl aminopeptidase
  • Aminopeptidase M
  • Aminopeptidase N
  • ANPEP
  • AP-M
  • APN
  • AP-N
  • CD13 antigen
  • CD13
  • CD13APN
  • EC 3.4.11
  • EC 3.4.11.2
  • gp150
  • LAP1
  • Microsomal aminopeptidase
  • Myeloid plasma membrane glycoprotein CD13
  • p150
  • PEPN
  • PEPNhAPN

Background

The human 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 human 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 (3).

  1. Olsen, J. et al. (1988) FEBS Lett. 238:307.
  2. Look, A.T. et al. (1989) J. Clin. Invest. 83:1299.
  3. Turner, A.J. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 289, Academic Press, San Diego.

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3815-ZN
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Publications for Aminopeptidase N/CD13 (3815-ZN)(5)

We have publications tested in 3 confirmed species: Human, Mouse, N/A.

We have publications tested in 3 applications: Bioassay, ELISA (Standard), Enzyme Assay.


Filter By Application
Bioassay
(3)
ELISA (Standard)
(1)
Enzyme Assay
(1)
All Applications
Filter By Species
Human
(3)
Mouse
(1)
N/A
(1)
All Species

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Bioinformatics

Gene Symbol ANPEP
Uniprot