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Recombinant Human Aminopeptidase N/CD13 His Avi Protein, CF

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Biotinylated Recombinant Human Aminopeptidase N/CD13 His-tag Avi-tag Protein (Catalog # AVI3815) binds Human Aminopeptidase N/CD13 Antibody (AF3815) with an ED50 of 1.80-18.0 ng/mL.
Biotinylated Recombinant Human Aminopeptidase N/CD13 His-tag Avi-tag Protein (Catalog # AVI3815) is measured by its ability to cleave the fluorogenic peptide substrate, Ala-7-amido-4-methylcoumarin (Ala-AMC).
2 μg/lane of Biotinylated Recombinant Human Aminopeptidase N/CD13 His-tag Avi-tag Protein (Catalog # AVI3815) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Binding Activity, Enzyme Activity
Format
Carrier-Free

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Recombinant Human Aminopeptidase N/CD13 His Avi Protein, CF Summary

Additional Information
Biotinylated, His-tag, Avi-tag
Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Ala-7-amido-4-methylcoumarin (Ala-AMC). The specific activity is >5000 pmol/min/μg, as measured under the described conditions. Measured by its binding ability in a functional ELISA. Biotinylated Recombinant Human Aminopeptidase N/CD13 His-tag Avi-tag (Catalog # AVI3815) binds Human Aminopeptidase N/CD13 Antibody (Catalog # AF3815) with an ED50 of 1.80-18.0 ng/mL.
Source
Chinese Hamster Ovary cell line, CHO-derived human Aminopeptidase N/CD13 protein
Human APN
(Lys69-Lys967)
Accession # AAA51719.1
6-His tagAvi-tag
N-terminusC-terminus
N-terminal Sequence
Lys69
Structure / Form

Biotinylated via Avi-tag

Protein/Peptide Type
Recombinant Enzymes
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Binding Activity2
  • Enzyme Activity
Theoretical MW
105 kD.
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
116-130 kDa, under 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 MES and NaCl.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 0.05%(w/v) Brij 35, pH 7.0
  • Biotinylated Recombinant Human Aminopeptidase N/CD13 His-tag Avi-tag (rhAPN/His/Avitag) (Catalog # AVI3815)
  • Substrate: Ala-AMC, 10 mM stock in DMSO
  • Black 96-well Plate
  • Plate Reader with Fluorescence Read Capability
  1. Dilute rhAPN/His/Avitag to 0.2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer.
  3. Load into a plate 50 µL of 0.2 µg/mL rhAPN/His/Avitag, and start the reaction by adding 50 µL of 400 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 400 µM 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.
Per Well:
  • rhAPN/His/Avitag: 0.01 µg
  • Substrate: 200 µM

Notes

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

Alternate Names for Recombinant Human Aminopeptidase N/CD13 His Avi Protein, CF

  • 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

Background

Aminopeptidase N (APN) is a type II transmembrane zinc-dependent metalloprotease that is widely expressed in many cells, tissues, and species. APN forms an active homodimer where each monomer contains a short cytoplasmic tail, a transmembrane region, and an extracellular domain with a Ser/Thr rich region and a metalloprotease catalytic domain (1,2). APN is also known as leukocyte surface differentiation antigen CD13, microsomal aminopeptidase, alanyl aminopeptidase, and aminopeptidase M (1-3). The protein cleaves the N-terminal neutral amino acids from bioactive peptides like neuropeptides and immunomodulatory peptides including oxytocin, vasopressin, angiotensin III, and tuftsin resulting in their degradation or modulation (2,4). Consequently, APN is implicated to play many biological roles within the immune system, angiogenesis, mestastasis, and cancer (2). Both APN expression and activity are altered in cancer making it a marker and a pharmacological intervention target (5,6). It is targeted in CAR T therapies for cancers such as B-cell lymphoma and multiple myeloma (2,7). In addition, APN serves as a receptor for several human viruses such as coronavirus 229E (8) and human cytomegalovirus making it of interest as a target in these fields (9,10).

  1. Look, A.T. et al. (1989) J. Clin. Invest. 83:1299.
  2. Lendeckel, U. et al. (2023) Biomedicines. 11:724.
  3. Olsen, J. et al. (1988) FEBS Lett. 238:307.
  4. Hansen, A.S. et al. (1993) Eur. J. Immunol. 23:2358.
  5. Li, H. et. al. (2019) Chem. Sci. 10:1619
  6. Shimizu, T. et. al. (2024) Front. Surg. 11:1298709.
  7. He, X. et. al. (2020) Blood 135:713.
  8. Yeager, C.L. et. al. (1992) Nature 357:420.
  9. Soderberg, C. et. al. (1993) J. Virol. 67: 6576
  10. Fu, Z. et. al. (2024) J. Virol. 98:e0123923.

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