Recombinant Human Aminopeptidase LRAP/ERAP2 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human Aminopeptidase LRAP/ERAP2 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate, Arg-7-amido-4-methylcoumarin (Arg-AMC). The specific activity is >50 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Aminopeptidase LRAP/ERAP2 protein
  Ala56-Thr960, with an N-terminal 6-His tag
• Accession #: Q6P179
• N-terminal Sequence: His
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ERAP2
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 105 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: 122 kDa, reducing conditions

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 Tris, NaCl and Glycerol.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 100 mM MES, pH 6.5
  • Recombinant Human Aminopeptidase LRAP/ERAP2 (Catalog # 3830-ZN)
  • Substrate: H-Arg-AMC (Bachem, Catalog # I-1050)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhLRAP to 5 µg/mL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. Load 50 µL of 5 µg/mL rhLRAP into a plate, and start the reaction by adding 50 µL of 100 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-Aldrich, Catalog # A9891). Per Well:

  • rhLRAP: 0.25 µg
  • Substrate: 50 µM

Notes

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

Alternate Names for Recombinant Human Aminopeptidase LRAP/ERAP2 Protein, CF

• Aminopeptidase LRAP
• EC 3.4.11
• EC 3.4.11.3
• endoplasmic reticulum aminopeptidase 2
• ERAP2
• FLJ23633
• FLJ23701
• FLJ23807
• Leukocyte-derived arginine aminopeptidase
• LRAPL-RAPEC 3.4.11.-

Background

Leukocyte-derived arginine aminopeptidase (LRAP) and endoplasmic reticulum-aminopeptidase 2 (ERAP2) are the two names given to the protein encoded by the LRAP gene (1, 2). Induced by interferon-gamma , LRAP is able to trim various MHC class I antigenic peptide precursors. It belongs to the oxytocinase subfamily of M1 aminopeptidases, which also includes aminopeptidases PILS/ARTS1/ERAP1 and LNPEP/PLAP (1, 3). In addition to antigen presentation, the members of this subfamily are also important in maintenance of pregnancy, memory retention, and blood pressure regulation (3).

1. Tanioka, T. et al. (2003) J. Biol. Chem. 278:32275.
2. Tanioka, T. et al. (2005) FEBS J. 272:916.
3. Tsujimoto, M. and A. Hattori (2005) Biochim. Biophys. Acta 1751:9.

Publications for Aminopeptidase LRAP/ERAP2 (3830-ZN)(1)

Publication using 3830-ZN

• K Textoris-T, C Cammann, P Henklein, E Topfstedt, F Ebstein, S Henze, J Liepe, F Zhao, D Schadendor, B Dahlmann, W Uckert, A Paschen, M Mishto, U Seifert ER-aminopeptidase 1 determines the processing and presentation of an immunotherapy-relevant melanoma epitope Eur. J. Immunol., 2019-11-27;0(0):. 2019-11-27 [PMID: 31729751] (Enzyme Assay, Human)

Bioinformatics

• Gene Symbol: ERAP2
• Uniprot:
  • Q6P179()