Human 5'-Nucleotidase/CD73 Antibody Summary
Trp27-Lys547
Accession # AAH65937
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human 5'-Nucleotidase/CD73 by Western Blot. Western blot shows lysates of U266 human myeloma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human 5'-Nucleotidase/CD73 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5795) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for 5'-Nucleotidase/CD73 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
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5'‑Nucleotidase/CD73 in A431 Human Cell Line. 5'-Nucleotidase/CD73 was detected in immersion fixed A431 human epithelial carcinoma cell line using Sheep Anti-Human 5'-Nucleotidase/CD73 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5795) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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5'‑Nucleotidase/CD73 in Human Cervical Cancer Tissue. 5'-Nucleotidase/CD73 was detected in immersion fixed paraffin-embedded sections of human cervical cancer tissue using Sheep Anti-Human 5'-Nucleotidase/CD73 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5795) at 1 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 plasma membranes in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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Detection of Human 5'‑Nucleotidase/CD73 by Simple WesternTM. Simple Western lane view shows lysates of U266 human myeloma cell line, loaded at 0.5 mg/mL. A specific band was detected for 5'-Nucleotidase/CD73 at approximately 86 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human 5'-Nucleotidase/CD73 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5795) 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.
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Detection of Human 5'‑Nucleotidase/CD73 by Simple WesternTM. Simple Western lane view shows lysates of U-87 MG human glioblastoma/astrocytoma cell line, U-251 MG human glioblastoma cell line, and A431 human epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for 5'-Nucleotidase/CD73 at approximately 86 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human 5'-Nucleotidase/CD73 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5795) 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. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
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Detection of 5'‑Nucleotidase/CD73 in Human Colon Cancer. Formalin-fixed paraffin-embedded tissue sections of human colon cancer were probed for NT5E mRNA (ACD RNAScope Probe, catalog #437931; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using sheep anti-human NT5E polyclonal antibody (R&D Systems catalog # AF5795) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-sheep IgG VisUCyte HRP Polymer Antibody (Catalog # VC006) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.
Reconstitution Calculator
Preparation and Storage
- 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.
Background: 5'-Nucleotidase/CD73
5'-Nucleotidase (also [ecto]-5'-nucleotidase/5'-NT, designated CD73) is a variably glycosylated, 69-73 kDa member of the 5'-Nucleotidase family of enzymes. It is expressed on multiple cell types, including vascular endothelium, transitional and nonkeratinized epithelium, cardiomyocytes, small intestine epithelium, FoxP3+ Treg lymphocytes, FDCs and B cells. 5'-Nucleotidase hydrolyzes AMP to adenosine and phosphate. This creates diffusible nucleosides necessary for cell homeostasis, and a ligand for cell membrane adenosine receptors. Mature human 5'-Nucleotidase is a 523 amino acid (aa) GPI-linked protein (aa 27-549). It contains a large
Zn‑dependent nucleotidase catalytic region (aa 28-532) and a C-terminal substrate binding site (aa 500-506). On the cell surface it exists as a disulfide-linked homodimer. Two splice variants are reported. One shows a deletion of aa 405-454, and a second possesses a 12 aa substitution for aa 253-574. Over aa 1-511, human 5'‑Nucleotidase shares 88% aa identity with both mouse and rat 5'-Nucleotidase.
Product Datasheets
Citations for Human 5'-Nucleotidase/CD73 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 2
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Selection of Highly Proliferative and Multipotent Meniscus Progenitors through Differential Adhesion to Fibronectin: A Novel Approach in Meniscus Tissue Engineering
Authors: JV Korpershoe, M Rikkers, TS de Windt, MA Tryfonidou, DBF Saris, LA Vonk
International Journal of Molecular Sciences, 2021-08-10;22(16):.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
NAD+ augmentation restores mitophagy and limits accelerated aging in Werner syndrome
Authors: EF Fang, Y Hou, S Lautrup, MB Jensen, B Yang, T SenGupta, D Caponio, R Khezri, TG Demarest, Y Aman, D Figueroa, M Morevati, HJ Lee, H Kato, H Kassahun, JH Lee, D Filippelli, MN Okur, A Mangerich, DL Croteau, Y Maezawa, CA Lyssiotis, J Tao, K Yokote, TE Rusten, MP Mattson, H Jasper, H Nilsen, VA Bohr
Nat Commun, 2019-11-21;10(1):5284.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
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