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Recombinant Human Aminopeptidase N/CD13 His Avi Protein, CF

Catalog #: AVI3815 Datasheet / COA / SDS
Biotinylated, His-tag, Avi-tag
Catalog # Availability Size / Price Qty
AVI3815-010
Biotinylated Recombinant Human Aminopeptidase N/CD13 His-tag Avi-tag Protein Binding Activity.
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 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 >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
Analysis
Lys69
Structure / Form

Biotinylated via Avi-tag

Predicted Molecular Mass
105 kD
SDS-PAGE
116-130 kDa, under reducing conditions.

Product Datasheets

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AVI3815

Product Datasheet
COA
SDS

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.

AVI3815

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, 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

Scientific Data

Binding Activity View Larger

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.

Enzyme Activity View Larger

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).

SDS-PAGE View Larger

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 Coomassie® Blue staining, showing bands at 116-130 kDa, under reducing conditions.

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

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).

References
  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.
Entrez Gene IDs
290 (Human); 16790 (Mouse); 397520 (Porcine); 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

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