Recombinant Human Aminopeptidase N/CD13 Protein, CF Summary
Lys69-Lys967, with a C-terminal 10-His tag
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.
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.
|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.
- 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
- Dilute rhCD13 to 0.2 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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
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).
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).
- Olsen, J. et al. (1988) FEBS Lett. 238:307.
- Look, A.T. et al. (1989) J. Clin. Invest. 83:1299.
- Turner, A.J. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 289, Academic Press, San Diego.
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.
Citations: Showing 1 - 6
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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):.
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance (SPR
Localization, Shedding, Regulation and Function of Aminopeptidase N/CD13 on Fibroblast like Synoviocytes
PLoS ONE, 2016;11(9):e0162008.
Sample Types: Plasma
Applications: ELISA (Standard)
The Aminopeptidase CD13 Induces Homotypic Aggregation in Neutrophils and Impairs Collagen Invasion
PLoS ONE, 2016;11(7):e0160108.
Sample Types: Whole Cells
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.
Sample Types: Whole Cells
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.
Sample Types: Whole Cells
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.
Sample Types: Peptide
Applications: Enzyme Assay
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