Recombinant Human Aminopeptidase P2/XPNPEP2 Protein, CF

Catalog # Availability Size / Price Qty
2490-ZN-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human Aminopeptidase P2/XPNPEP2 Protein, CF Summary

Product Specifications

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, H-Lys(2-Aminobenzoyl)-Pro-Pro-p-Nitroanilide (K(Abz)PP-pNA). The specific activity is >300 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Aminopeptidase P2/XPNPEP2 protein
His22-Ala650, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
His22 & Lys24
Predicted Molecular Mass
72 kDa
SDS-PAGE
85 kDa, reducing conditions

Product Datasheets

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2490-ZN

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.

2490-ZN

Formulation Supplied as a 0.2 μm filtered solution in Tris 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: 100 mM HEPES, 0.5 mM MnCl2, pH 8.0
  • Recombinant Human Aminopeptidase P2/XPNPEP2 (rhXPNPEP2) (Catalog # 2490-ZN)
  • Substrate: Lys(Abz)-Pro-Pro-pNA (Bachem, Catalog # L-1980), 10 mM stock in deionized water
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhXPNPEP2 to 1 ng/µL in Assay Buffer. Incubate at room temperature for 30 minutes.
  2. Dilute Substrate to 100 μM in Assay Buffer.
  3. Load into plate 50 μL of 1 ng/µL of rhXPNPEP2 and start the reaction by adding 50 µL of 100 µM Substrate.  Include a Substrate Blank of 50 μL Assay Buffer and 50 µL of 100 µM Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 410 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 ABZ-Gly (Bachem, Catalog # E-2920).

Per Well:
  • rhXPNPEP2: 0.05 µg
  • Substrate: 50 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 P2/XPNPEP2

The human XPNPEP2 gene encodes Aminopeptidase P2 (APP2), which is also known as X-prolyl Aminopeptidase 2 or membrane bound Aminopeptidase P (1‑4). It is a member of the M24 family of metalloproteases, which also contains methionine Aminopeptidases, X-Pro dipeptidase, Aminopeptidase P1, Aminopeptidase P homolog, proliferation-associated protein 1, and suppressor of Ty homolog or chromatin-specific transcription elongation factor large subunit (5). Mammalian APP2 are predicted to be GPI-anchored membrane proteases and their biological functions have been reviewed (6). Human APP2 is widely expressed in many adult tissues with the highest levels in the kidney (7). The purified recombinant human APP2 corresponds to the ectodomain and is shown here to be an active aminopeptidase, removing a N-terminal amino acid from a peptide that contains a Pro residue at the second position.

References
  1. Venema, R.C. et al. (1997) Biochim. Biophys. Acta 1354:45.
  2. Sprinkle, T.J. et al. (1998) Genomics 50:114.
  3. Cottrell, G.S. et al. (1998) Biochem. Soc. Trans. 26:S248.
  4. Prueitt, R.L. et al. (2000) Cytogenet. Cell Genet. 89:44.
  5. Barrett, A.J. et al. (2004) Handbook of Proteolytic Enzymes, Elsevier Academic Press, San Diego.
  6. Simmons, W.H. (2004) in Handbook of Proteolytic Enzymes (Barrett, A.J. et al. eds.) p. 934, Elsevier Academic Press, San Diego.
  7. Ersahin C. et al. (2005) Arch. Biochem. Biophys. 435:303.
Entrez Gene IDs
7512 (Human); 170745 (Mouse)
Alternate Names
Aminoacylproline aminopeptidase; Aminopeptidase P2; APP2; EC 3.4.11.9; mAmP; Membrane-bound aminopeptidase P; Membrane-bound AmP; Membrane-bound APP; xaa-Pro aminopeptidase 2; XPNPEP2; X-Pro aminopeptidase 2; X-prolyl aminopeptidase (aminopeptidase P) 2, membrane-bound

Product Specific Notices

The purchase of this product conveys to the buyer the limited, non-exclusive, non-transferable right (without the right to resell, repackage, or further sublicense) to use these reagents for non-commercial research purposes only. No other license is granted to the buyer whether expressly, by implication, by estoppel or otherwise. In particular, the purchase of this product does not include nor carry any right or license to use, develop, or otherwise exploit this product commercially, which includes without limitation, provision of services to a third party, generation of commercial databases, clinical diagnostics or therapeutics, or drug development. This Product is manufactured under a license to U.S. Patent Nos. 6,399,349 and 7,273,718.

Any party desiring rights under this patent should contact Ryogen LLC, Montebello Park, 75 Montebello Road, Suffern, NY 10901.

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