Recombinant Mouse Aminopeptidase PILS/ARTS1 Protein, CF

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

Recombinant Mouse Aminopeptidase PILS/ARTS1 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate, Leu-AMC. The specific activity is >100 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse Aminopeptidase PILS/ARTS1 protein
  Ser27-Leu930, with a C-terminal 10-His tag
• Accession #: Q6GTP5
• N-terminal Sequence: Ser27
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Erap1
• Purity: >95%, 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: 111 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: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 25 mM Tris, pH 8.0
  • Recombinant Mouse Aminopeptidase PILS/ARTS1 (rmARTS1) (Catalog # 2500-ZN)
  • Substrate: Leu-AMC (Bachem, Catalog # I-1240)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmARTS1 to 2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load 50 µL of the 2 µg/mL rmARTS1 into a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, 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 AMC (Sigma, Catalog # A-9891).

  Per Well:
  • rmARTS1: 0.1 µg
  • Substrate: 100 µM

Notes

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

Alternate Names for Recombinant Mouse Aminopeptidase PILS/ARTS1 Protein, CF

• Adipocyte-derived leucine aminopeptidase
• A-LAP
• A-LAPALAP
• Aminopeptidase PILS
• ARTS1
• ARTS1aminopeptidase regulator of TNFR1 shedding
• ARTS-1PILSAP
• EC 3.4.11
• EC 3.4.11.-
• EC 3.4.11.1
• endoplasmic reticulum aminopeptidase 1
• endoplasmic reticulum aminopeptidase associated with antigen processing
• ERAAP1
• ERAP1
• KIAA0525APPILS
• PILS-AP
• PILS-APERAAP
• Puromycin-insensitive leucyl-specific aminopeptidase
• Type 1 tumor necrosis factor receptor shedding aminopeptidase regulator

Background

The mouse ARTS1 gene encodes aminopeptidase PILS (Puromycin-Insensitive Leucyl-Specific), which is also known as adipocyte-derived leucine aminopeptidase, type 1 tumor necrosis factor receptor shedding aminopeptidase regulator and ERAAP (the aminopeptidase associated with antigen processing in the endoplasmic reticulum) (1-3). The deduced amino acid sequence of mouse ARTS1 consists of a signal peptide and a large ectodomain. Widely expressed and releasing an N‑terminal amino acid (Leu), the enzyme may play a role in many processes such as antigen processing and angiogenesis.

1. Miyashita, H. et al. (2002) Blood 99:3241.
2. Serwold, T. et al. (2002) Nature 419:480.
3. Schomburg, L. (2004) in Handbook of Proteolytic Enzymes. Barrett, et al. eds. p. 311, Academic Press, San Diego.

Publications for Aminopeptidase PILS/ARTS1 (2500-ZN)(1)

Publication using 2500-ZN

• Infantes S, Samino Y, Lorente E, Jimenez M, Garcia R, Del Val M, Lopez D H-2Ld class I molecule protects an HIV N-extended epitope from in vitro trimming by endoplasmic reticulum aminopeptidase associated with antigen processing. J. Immunol., 2010-03-03;184(7):3351-5. 2010-03-03 [PMID: 20200278] (Enzyme Assay, Mouse)

Bioinformatics

• Gene Symbol: Erap1
• Uniprot:
  • Q6GTP5()