Recombinant Human Aminopeptidase N/CD13 Protein, CF

Catalog # Availability Size / Price Qty
3815-ZN-010
Recombinant Human Aminopeptidase N/CD13 Protein, CF Enzyme Activity
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Product Details
Citations (6)
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Recombinant Human Aminopeptidase N/CD13 Protein, CF Summary

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.
Activity
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
Analysis
Lys69
Predicted Molecular Mass
104 kDa
SDS-PAGE
131 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

3815-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.

Assay Procedure

Materials
  • 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

Data Image

Enzyme Activity View Larger

Recombinant Human Aminopeptidase N (Catalog # 3815-ZN) is measured by its ability to cleave the fluorogenic peptide substrate, Ala-7-amido-4-methylcoumarin (Ala-AMC).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Aminopeptidase N/CD13

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).

References
  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.
Entrez Gene IDs
290 (Human); 16790 (Mouse); 102115663 (Cynomolgus Monkey)
Alternate Names
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

Citations for Recombinant Human Aminopeptidase N/CD13 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.

6 Citations: Showing 1 - 6
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  1. A human antithrombin isoform dampens inflammatory responses and protects from organ damage during bacterial infection
    Authors: P Papareddy, M Rossnagel, F Doreen Hol, G Kilic, S Veerla, C Naudin, E Smeds, J Westman, I Martinez-M, A Egesten, ME de la More, J Corral, A Linder, A Artoni, M Abbattista, C Novembrino, C Herbert Br, I Martinelli, G Kasetty, H Herwald
    Nat Microbiol, 2019;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Surface Plasmon Resonance (SPR
  2. Localization, Shedding, Regulation and Function of Aminopeptidase N/CD13 on Fibroblast like Synoviocytes
    PLoS ONE, 2016;11(9):e0162008.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA (Standard)
  3. The Aminopeptidase CD13 Induces Homotypic Aggregation in Neutrophils and Impairs Collagen Invasion
    PLoS ONE, 2016;11(7):e0160108.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Cot/tpl2 participates in the activation of macrophages by adiponectin.
    Authors: Sanz-Garcia C, Nagy L, Lasuncion M, Fernandez M, Alemany S
    J Leukoc Biol, 2014;95(6):917-30.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  5. Adiponectin modulates NK-cell function.
    Authors: Wilk S, Jenke A, Stehr J, Yang C, Bauer S, Goldner K, Kotsch K, Volk H, Poller W, Schultheiss H, Skurk C, Scheibenbogen C
    Eur J Immunol, 2013;43(4):1024-33.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  6. Cysteine cathepsins S and L modulate anti-angiogenic activities of human endostatin.
    Authors: Veillard F, Saidi A, Burden RE, Scott CJ, Gillet L, Lecaille F, Lalmanach G
    J. Biol. Chem., 2011;286(43):37158-67.
    Species: N/A
    Sample Types: Peptide
    Applications: Enzyme Assay

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