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Human 5'‑Nucleotidase/CD73 Antibody
Catalog # AF5795 | R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
Species Reactivity
Human
Applications
Immunocytochemistry, Immunohistochemistry, Simple Western, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
Product Summary for Human 5'‑Nucleotidase/CD73 Antibody
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human 5'‑Nucleotidase/CD73
Trp27-Lys547
Accession # AAH65937
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.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human 5'‑Nucleotidase/CD73 Antibody
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.
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.
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.Applications for Human 5'‑Nucleotidase/CD73 Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line
Sample: Immersion fixed A431 human epithelial carcinoma cell line
Immunohistochemistry
1-15 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
10 µg/mL
Sample: U266 human myeloma cell line, U‑87 MG human glioblastoma/astrocytoma cell line, U‑251 MG human glioblastoma cell line, and A431 human epithelial carcinoma cell line
Sample: U266 human myeloma cell line, U‑87 MG human glioblastoma/astrocytoma cell line, U‑251 MG human glioblastoma cell line, and A431 human epithelial carcinoma cell line
Western Blot
1 µg/mL
Sample: U266 human myeloma cell line
Sample: U266 human myeloma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 1 review rated 5 using AF5795 in the following applications:
Published Applications
Read 2 publications using AF5795 in the following applications:
Formulation, Preparation and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
Product Documents for Human 5'‑Nucleotidase/CD73 Antibody
Product Specific Notices for Human 5'‑Nucleotidase/CD73 Antibody
For research use only
Citations for Human 5'‑Nucleotidase/CD73 Antibody (2)
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Citations are publications that use Bio-Techne products. Selected citations for Human 5'‑Nucleotidase/CD73 Antibody include:
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Species: Human
Sample Types: Whole Cells
Applications: Flow CytometryJV Korpershoe et al. (2021-08-10), Selection of Highly Proliferative and Multipotent Meniscus Progenitors through Differential Adhesion to Fibronectin: A Novel Approach in Meniscus Tissue Engineering International Journal of Molecular Sciences, 2021-022(16).
PMID: 34445320 -
Species: Human
Sample Types: Cell Lysates
Applications: Western BlotEF Fang et al. (2019-11-21), NAD+ augmentation restores mitophagy and limits accelerated aging in Werner syndrome Nat Commun, 2019-110(1):5284.
PMID: 31754102
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Customer Reviews for Human 5'‑Nucleotidase/CD73 Antibody (1)
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1 customer
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Application: immunofluorescence - paraffinSample Tested: human tonsilSpecies: HumanVerified Customer | Posted 05/02/2019pH 9 heat-induced antigen retrieval
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FAQs for Human 5'‑Nucleotidase/CD73 Antibody
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Q: We are interested in CD73 antibodies that could react with dog, please let me know which products in your catalog you would recommend.
A: 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.
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