5'-Nucleotidase/CD73 Antibody [Unconjugated]

Western blot shows lysates of U266 human myeloma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human5'‑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.

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.

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-Human5'‑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 the12-230 kDa separation system.

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.

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.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

5'-Nucleotidase/CD73 Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human 5'‑Nucleotidase/CD73
  Trp27-Lys547
  Accession # AAH65937
• Specificity: Detects human 5'‑Nucleotidase/CD73 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross‑reactivity with recombinant mouse CD73 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: NT5E

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Simple Western 10 ug/mL
  • Immunohistochemistry 1-15 ug/mL
  • Immunocytochemistry 5-15 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 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 5'-Nucleotidase/CD73 Antibody [Unconjugated]

• 5' nucleotidase (CD73)
• 5'-NT
• 5-NT
• 5'-Nucleotidase
• 5'-nucleotidase, ecto (CD73)
• CD73 antigen
• CD73
• E5NT
• EC 3.1.3.5
• ecto-5'-nucleotidase
• eN
• eNT
• NT
• NT55'-nucleotidase
• NT5E
• NTE
• Purine 5-Prime-Nucleotidase

Background

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.

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 5'-Nucleotidase/CD73 Antibody (AF5795)(1)

Publication using AF5795

• 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 NAD+ augmentation restores mitophagy and limits accelerated aging in Werner syndrome Nat Commun, 2019;10(1):5284. 2019 [PMID: 31754102] (Western Blot, Human)

Review for 5'-Nucleotidase/CD73 Antibody (AF5795) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous Human 05/02/2019

Summary

• Sample Tested: human tonsil
• Species: Human
• Lot: I don't have it any more

Comments

• Comments: pH 9 heat-induced antigen retrieval

FAQs for 5'-Nucleotidase/CD73 Antibody (AF5795). (Showing 1 - 1 of 1 FAQ).

1. We are interested in CD73 antibodies that could react with dog, please let me know which products in your catalog you would recommend.
   • We do not currently carry any CD73 antibodies that have been validated for use in dog samples. However, if you are interested in testing any of our CD73 antibodies with dog samples you would be eligible for our Innovators Reward Program, in which you can get a discount voucher for the purchase price of the product. Please contact us at innovators@novusbio.com with any questions regarding this program.

Bioinformatics

• Gene Symbol: NT5E
• Uniprot:
  • AAH65937()