Aminopeptidase N (APN/CD13) Activity Assay Kit (Fluorometric) (ab273292)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Fluorescent
- Platform: Microplate
- Sample type: Adherent cells, Cell Lysate, Plasma, Serum, Suspension cells, Tissue, Tissue Lysate
- Sensitivity: 100 µU
Overview
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Product name
Aminopeptidase N (APN/CD13) Activity Assay Kit (Fluorometric)
See all CD13 kits -
Detection method
Fluorescent -
Sample type
Serum, Plasma, Tissue, Adherent cells, Suspension cells, Cell Lysate, Tissue Lysate -
Assay type
Enzyme activity (quantitative) -
Sensitivity
100 µU -
Product overview
Aminopeptidase N (APN/CD13) Activity Assay Kit (Fluorometric) (ab273292) enables rapid measurement of APN activity, utilizing a fluorogenic substrate that is converted into a highly fluorescent product (Ex/Em = 384/502 nm). A selective APN inhibitor is provided for verification of specific activity.
The assay is simple, high-throughput adaptable and can detect a minimum of 100 μU APN activity in a variety of biological samples.
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Platform
Microplate
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests AFC Standard (1 mM) 1 x 100µl Aminopeptidase Assay Buffer 1 x 50ml Aminopeptidase N Inhibitor (Lyophilized) 1 vial Aminopeptidase N Positive Control (Lyophilized) 1 vial Aminopeptidase N Substrate (Lyophilized) 1 vial -
Research areas
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Function
Broad specificity aminopeptidase. Plays a role in the final digestion of peptides generated from hydrolysis of proteins by gastric and pancreatic proteases. May play a critical role in the pathogenesis of cholesterol gallstone disease. May be involved in the metabolism of regulatory peptides of diverse cell types including small intestinal and tubular epithelial cells, macrophages, granulocytes and synaptic membranes from the CNS. Found to cleave antigen peptides bound to major histocompatibility complex class II molecules of presenting cells and to degrade neurotransmitters at synaptic junctions. Is also implicated as a regulator of IL-8 bioavailability in the endometrium, and therefore may contribute to the regulation of angiogenesis. Is used as a marker for acute myeloid leukemia and plays a role in tumor invasion. In case of human coronavirus 229E (HCoV-229E) infection, serves as receptor for HCoV-229E spike glycoprotein. Mediates as well human cytomegalovirus (HCMV) infection. -
Tissue specificity
Expressed in epithelial cells of the kidney, intestine, and respiratory tract; granulocytes, monocytes, fibroblasts, endothelial cells, cerebral pericytes at the blood-brain barrier, synaptic membranes of cells in the CNS. Also expressed in endometrial stromal cells, but not in the endometrial glandular cells. Found in the vasculature of tissues that undergo angiogenesis and in malignant gliomas and lymph node metastases from multiple tumor types but not in blood vessels of normal tissues. A soluble form has been found in plasma. It is found to be elevated in plasma and effusions of cancer patients. -
Sequence similarities
Belongs to the peptidase M1 family. -
Domain
Amino acids 260-353 are essential to mediate susceptibility to infection with HCoV-229E (in porcine/human chimeric studies) and more specifically amino acids 288-295 (mutagenesis studies). -
Post-translational
modificationsSulfated.
N- and O-glycosylated.
May undergo proteolysis and give rise to a soluble form. -
Cellular localization
Cell membrane. Cytoplasm > cytosol. A soluble form has also been detected. - Information by UniProt
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Alternative names
- Alanyl (membrane) aminopeptidase
- Alanyl aminopeptidase
- Aminopeptidase M
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Images
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Typical standard curve – data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.
Standard curve of 7-amino-4-trifluoromethylcoumarin (AFC) fluorescence. One mole of AFC corresponds to the metabolism of one mole of Aminopeptidase N Substrate).
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Kinetics of APN Substrate metabolism in rat kidney lysate (5 µg protein), HT-1080 cell and HepG2 cell lysates (each 10 µg protein), as well as pooled normal human serum and serum from breast cancer patients (each 10 µl undiluted serum) in the presence and absence of the included selective APN inhibitor.
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Quantification of APN activity in tissue/cell lysates and human serum samples (mean ± SEM at least 3 replicates). Assays were performed according to the kit protocol.