Recombinant Mouse Aminopeptidase N/CD13 Protein, CF

Catalog # Availability Size / Price Qty
2335-ZN-010
Product Details
Citations (3)
FAQs
Reviews

Recombinant Mouse 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 >1,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse Aminopeptidase N/CD13 protein
Lys69-Ser966, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Lys69
Predicted Molecular Mass
104 kDa
SDS-PAGE
133 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.

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.

Assay Procedure

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
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 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.
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 Mouse 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.

3 Citations: Showing 1 - 3
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  1. Adiponectin attenuates profibrotic extracellular matrix remodeling following cardiac injury by up-regulating matrix metalloproteinase 9 expression in mice
    Authors: A Jenke, R Schur, C Röger, Z Karadeniz, M Grüger, L Holzhauser, K Savvatis, W Poller, HP Schultheis, U Landmesser, C Skurk
    Physiol Rep, 2017;5(24):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  2. 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: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  3. The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) interacts with membrane type 1 matrix metalloproteinase and CD13/aminopeptidase N and modulates their endocytic pathways.
    Authors: Miki T, Takegami Y, Okawa K, Muraguchi T, Noda M, Takahashi C
    J. Biol. Chem., 2007;282(16):12341-52.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay

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