Human/Mouse/Rat 5'-Nucleotidase/CD73 Antibody Summary
Accession # P21589
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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.
Detection of Human, Mouse, and Rat 5'‑Nucleotidase/CD73 by Western Blot. Western blot shows lysates of HT-29 human colon adenocarcinoma cell line, Huh-7 human hepatoma cell line, U266 human myeloma cell line, mouse brain tissue, and rat brain tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat 5'-Nucleotidase/CD73 Monoclonal Antibody (Catalog # MAB57951) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 1.
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 Mouse Anti-Human/Mouse/Rat 5'-Nucleotidase/CD73 Monoclonal Antibody (Catalog # MAB57951) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and cell membranes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Human 5'‑Nucleotidase/CD73 by Simple WesternTM. Simple Western lane view shows lysates of 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 20 µg/mL of Mouse Anti-Human/Mouse/Rat 5'‑Nucleotidase/CD73 Monoclonal Antibody (Catalog # MAB57951). 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.
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.
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.
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