Human/Mouse/Rat 5'-Nucleotidase/CD73 Antibody
R&D Systems, part of Bio-Techne | Catalog # MAB57951
Key Product Details
Species Reactivity
Human, Mouse, Rat
Applications
Immunohistochemistry, Simple Western, Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2B Clone # 606117
Product Specifications
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human 5'‑Nucleotidase/CD73
Trp27-Ile511
Accession # P21589
Trp27-Ile511
Accession # P21589
Specificity
Detects human 5'-Nucleotidase/CD73 in direct ELISAs. Detects human, mouse, and rat 5'-Nucleotidase/CD73 in Western blots.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
Scientific Data Images for Human/Mouse/Rat 5'-Nucleotidase/CD73 Antibody
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.Applications for Human/Mouse/Rat 5'-Nucleotidase/CD73 Antibody
Application
Recommended Usage
Immunohistochemistry
1-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human cervical cancer tissue
Sample: Immersion fixed paraffin-embedded sections of human cervical cancer tissue
Simple Western
20 µg/mL
Sample: U‑251 MG human glioblastoma cell line and A431 human epithelial carcinoma cell line
Sample: U‑251 MG human glioblastoma cell line and A431 human epithelial carcinoma cell line
Western Blot
2 µg/mL
Sample: 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
Sample: 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
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & 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.
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.
Alternate Names
5-NT, CD73, E5NT, eNT, NT5E, NTE
Gene Symbol
NT5E
UniProt
Additional 5'-Nucleotidase/CD73 Products
- All Products for 5'-Nucleotidase/CD73
- 5'-Nucleotidase/CD73 Activity Assays
- 5'-Nucleotidase/CD73 cDNA Clones
- 5'-Nucleotidase/CD73 ELISA Kits
- 5'-Nucleotidase/CD73 Lysates
- 5'-Nucleotidase/CD73 Primary Antibodies
- 5'-Nucleotidase/CD73 Proteins and Enzymes
- 5'-Nucleotidase/CD73 Small Molecules and Peptides
Product Documents for Human/Mouse/Rat 5'-Nucleotidase/CD73 Antibody
Product Specific Notices for Human/Mouse/Rat 5'-Nucleotidase/CD73 Antibody
For research use only
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