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Anti-CD73 (extracellular) Antibody

Ecto-5-nucleotidase, E5NT, 5'-Nucleotidase, 5'-NT, NT5E

Cat #: ANT-066
Alternative Name Ecto-5-nucleotidase, E5NT, 5'-Nucleotidase, 5'-NT, NT5E
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)KGSTLKKAFEHSVHR, corresponding to amino acid residues 429-433 of mouse CD73 (Accession Q61503). Extracellular, N-terminus.
Accession (Uniprot) Number Q61503
Gene ID 23959
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Human, rat - identical.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ifc, ih, wb
May also work in: ic*, ip*, lci*
Western blot
  • Western blot analysis of mouse brain membranes (lanes 1 and 3) and rat brain lysate (lanes 2 and 4):
    Western blot analysis of mouse brain membranes (lanes 1 and 3) and rat brain lysate (lanes 2 and 4):
    1,2. Anti-CD73 (extracellular) Antibody (#ANT-066), (1:200).
    3,4. Anti-CD73 (extracellular) Antibody, preincubated with CD73 (extracellular) Blocking Peptide (#BLP-NT066).
  • Western blot analysis of human U-87 MG glioblastoma cell line lysate (lanes 1 and 3) and human MDA-MB-231 breast adenocarcinoma cell line lysate (lanes 2 and 4):
    Western blot analysis of human U-87 MG glioblastoma cell line lysate (lanes 1 and 3) and human MDA-MB-231 breast adenocarcinoma cell line lysate (lanes 2 and 4):
    1,2. Anti-CD73 (extracellular) Antibody (#ANT-066), (1:200).
    3,4. Anti-CD73 (extracellular) Antibody, preincubated with CD73 (extracellular) Blocking Peptide (#BLP-NT066).
Immunohistochemistry
  • Expression of CD73 in mouse striatum and hippocampus.
    Expression of CD73 in mouse striatum and hippocampus.
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections with Anti-CD73 (extracellular) Antibody (#ANT-066), (1:200), followed by goat anti-rabbit-AlexaFluor-488. A. CD73 staining (green) in mouse striatum is detected in striatal cells (arrows). B. CD73 staining (green) in hippocampal CA1 region is detected in cells of the pyramidal layer (vertical arrow) and their apical dendrites (horizontal arrows). Cell nuclei are stained with DAPI (blue).
Indirect flow cytometry
  • Cell surface detection of CD73 in live intact human Jurkat T-cell leukemia cells:
    Cell surface detection of CD73 in live intact human Jurkat T-cell leukemia cells:
    ___ Cells.
    ___ Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-CD73 (extracellular) Antibody (#ANT-066), (5 µg) + goat-anti-rabbit-FITC.
References
  1. Chen, S. et al. (2019) Nat. Commun. 10, 150.
  2. Kordaß, T. et al. (2018) Front. Immunol. 9, 813.
  3. Wang, R. et al. (2017) Oncotarget 8, 57327.
Scientific background

CD73 (ecto-5′-nucleotidase, NT5E) is a glycosylphosphatidylinositol enzyme expressed on the cell surface of various cell types. CD73 is responsible for hydrolyzing extracellular adenosine monophosphate to adenosine and inorganic phosphate, which results in activation of the G-protein coupled receptors, namely A2AR and A2BR, to exert potent immunoregulatory activity.

CD73 plays an important role in homeostatic functions in healthy organs and tissues. The protein is involved as an inhibitory immune checkpoint molecule, it promotes immune escape of tumor cells since free adenosine generated by the protein inhibits cellular immune responses1,2.

CD73 primarily detected in cancer cells and in immune cells such as CD4+Foxp3+ regulatory T cells, and myeloid-derived suppressor cells that are recruited by the tumor2.

Studies show that CD73 expression is significantly up-regulated in numerous malignant tumors such as breast cancer, colorectal cancer, prostate cancer, ovarian cancer, and gallbladder cancer3.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 08/06/2021

Anti-CD73 (extracellular) Antibody (#ANT-066) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry, and live cell flow cytometry applications. It has been designed to recognize NT5E from human, rat and mouse samples.

For research purposes only, not for human use
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