Aminopeptidase N/CD13 Antibody [Unconjugated]

HAF016). A specific band was detected for Aminopeptidase N/CD13 at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." > HAF016). A specific band was detected for Aminopeptidase N/CD13 at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." title="Western blot shows lysates of human kidney tissue and human prostate tissue. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Aminopeptidase N/CD13 at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." />

Western blot shows lysates of human kidney tissue and human prostate tissue. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # <a class=

5-001-A, open histogram), followed by APC-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). View our protocol for Staining Membrane-associated Proteins." > 5-001-A, open histogram), followed by APC-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). View our protocol for Staining Membrane-associated Proteins." title="U937 human histiocytic lymphoma cell line was stained with Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by APC-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). View our protocol for Staining Membrane-associated Proteins." />

U937 human histiocytic lymphoma cell line was stained with Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815, filled histogram) or isotype control antibody (Catalog # <a class=

CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." > CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." title="Aminopeptidase N/CD13 was detected in immersion fixed paraffin-embedded sections of human kidney using Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) at 0.3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." />

Aminopeptidase N/CD13 was detected in immersion fixed paraffin-embedded sections of human kidney using Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) at 0.3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # <a class=

HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." > HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." title="Simple Western lane view shows lysates of human small intestine tissue and human prostate tissue, loaded at 0.5 mg/mL. A specific band was detected for Aminopeptidase N/CD13 at approximately 177-198 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." />

Simple Western lane view shows lysates of human small intestine tissue and human prostate tissue, loaded at 0.5 mg/mL. A specific band was detected for Aminopeptidase N/CD13 at approximately 177-198 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # <a class=

HAF016). A specific band was detected for Aminopeptidase N/CD13 at approximately 150 kDa (as indicated) in the parental U937 human histiocytic lymphoma cell line, but is not detectable in knockout U937 human histiocytic lymphoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1." > HAF016). A specific band was detected for Aminopeptidase N/CD13 at approximately 150 kDa (as indicated) in the parental U937 human histiocytic lymphoma cell line, but is not detectable in knockout U937 human histiocytic lymphoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1." title="Western blot shows lysates of U937 human histiocytic lymphoma cell line and human APN knockout U937 human histiocytic lymphoma cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Aminopeptidase N/CD13 at approximately 150 kDa (as indicated) in the parental U937 human histiocytic lymphoma cell line, but is not detectable in knockout U937 human histiocytic lymphoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1." />

Western blot shows lysates of U937 human histiocytic lymphoma cell line and human APN knockout U937 human histiocytic lymphoma cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3815) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (<a class=

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, IP, CyTOF-ready, KO
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Aminopeptidase N/CD13 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Aminopeptidase N/CD13
  Lys69-Lys967
  Accession # P15144
• Specificity: Detects human Aminopeptidase N/CD13 in the direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 20% cross-reactivity with recombinant mouse APN is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: ANPEP
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 0.3-15 ug/mL
  • Immunoprecipitation 25 ug/mL
  • Knockout Validated 0.5 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for Aminopeptidase N/CD13 Antibody [Unconjugated]

• 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

The human ANPEP gene encodes aminopeptidase N (APN), which is also known as microsomal aminopeptiase, alanyl aminopeptidase, aminopeptidase M, CD13, or membrane protein p161 (1‑3). The deduced amino acid sequence of human APN consists of a short cytoplasmic tail (residues 2 to 8), a transmembrane region (residue 9 to 32), a Ser/Thr rich region and a zinc metalloprotease domain (residues 69 to 966). Widely expressed in many cells, tissues and species, APN cleaves the N-terminal amino acids from bioactive peptides, leading to their inactivation or degradation. The roles of APN in many fields, such as neuroscience, hematopoeitic cells, immune system, angiogenesis, cancer and viral infection, have been reviewed (3).

1. Olsen, J. et al. (1988) FEBS Lett. 238:307.
2. Look, A.T. et al. (1989) J. Clin. Invest. 83:1299.
3. Turner, A.J. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 289, Academic Press, San Diego.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Aminopeptidase N/CD13 Antibody (AF3815)(6)

Publications using AF3815

• Yongmei Liu, Danying Chen, Yuanyuan Wang, Xinglin Li, Yaruo Qiu, Mei Zheng et al. Characterization of CCoV-HuPn-2018 spike protein-mediated viral entry J Virol 9/28/2023 [PMID: 37676001]
• Minyong Chen, Diego M. Assis, Matthieu Benet, Colleen M. McClung, Elizabeth A. Gordon, Shourjo Ghose et al. Comparative site-specific N-glycoproteome analysis reveals aberrant N-glycosylation and gives insights into mannose-6-phosphate pathway in cancer Communications Biology 1/13/2023 [PMID: 36639722]
• Minyong Chen, Diego M. Assis, Matthieu Benet, Colleen M. McClung, Elizabeth A. Gordon, Shourjo Ghose et al. Comparative site-specific N-glycoproteome analysis reveals aberrant N-glycosylation and gives insights into mannose-6-phosphate pathway in cancer Communications Biology 2023-01-13 [PMID: 36639722] (Western Blot, Human)
• Lisa L Kirkemo, Susanna K Elledge, Jiuling Yang, James R Byrnes, Jeff E Glasgow, Robert Blelloch et al. Cell-surface tethered promiscuous biotinylators enable comparative small-scale surface proteomic analysis of human extracellular vesicles and cells eLife 3/8/2022 [PMID: 35257663]
• Bertram S, Dijkman R, Habjan M, Heurich A, Gierer S, Glowacka I, Welsch K, Winkler M, Schneider H, Hofmann-Winkler H, Thiel V, Pohlmann S TMPRSS2 activates the human coronavirus 229E for cathepsin-independent host cell entry and is expressed in viral target cells in the respiratory epithelium. J Virol, 2013-03-27;87(11):6150-60. 2013-03-27 [PMID: 23536651] (IHC, IHC-Fr, Human)
• Dijkman R, Jebbink M, Koekkoek S, Deijs M, Jonsdottir H, Molenkamp R, Ieven M, Goossens H, Thiel V, van der Hoek L Isolation and characterization of current human coronavirus strains in primary human epithelial cell cultures reveal differences in target cell tropism. J Virol, 2013-02-20;87(11):6081-90. 2013-02-20 [PMID: 23427150] (IHC, Human)

Bioinformatics

• Gene Symbol: ANPEP
• Uniprot:
  • P15144()