Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Dual RNAscope ISH-IHC Compatible, Immunocytochemistry, Simple Western

Cited:

Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all 5'-Nucleotidase/CD73 Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human 5'‑Nucleotidase/CD73 Antibody

Detection of Human 5'-Nucleotidase/CD73 antibody by Western Blot.

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.

Detection of Human 5'-Nucleotidase/CD73 antibody by Simple WesternTM.

Detection of Human 5'‑Nucleotidase/CD73 by Simple WesternTM.

Simple Western shows lysates of Exosome Standards (HT‑29) (NBP3-11685), Exosome Standards (U‑87 MG) (NBP2-49844) and A431 human epithelial carcinoma 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-Human 5'‑Nucleotidase/CD73 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5795). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

5'-Nucleotidase/CD73 antibody in A431 Human Cell Line by Immunocytochemistry (ICC).

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 antibody in Human Cervical Cancer Tissue by Immunohistochemistry (IHC-P).

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.

Detection of Human 5'-Nucleotidase/CD73 antibody by Simple WesternTM.

Detection of Human 5'‑Nucleotidase/CD73 by Simple WesternTM.

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.

Detection of 5'‑Nucleotidase/CD73 in Human Colon Cancer.

Formalin-fixed paraffin-embedded tissue sections of human colon cancer were probed for NT5E mRNA (ACD RNAScope Probe, catalog #437931; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using sheep anti-human NT5E polyclonal antibody (R&D Systems catalog # AF5795) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-sheep IgG VisUCyte HRP Polymer Antibody (Catalog # VC006) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Applications for Human 5'‑Nucleotidase/CD73 Antibody

Application
Recommended Usage

Dual RNAscope ISH-IHC Compatible

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer

Immunocytochemistry

5-15 µg/mL
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

Simple Western

10 µg/mL
Sample: Exosome Standards (HT-29) (Catalog # NBP3-11685), Exosome Standards (U-87 MG) (Catalog # NBP2-49844) and A431 human epithelial carcinoma cell line

Western Blot

1 µg/mL
Sample: U266 human myeloma cell line

Reviewed Applications

Read 1 review rated 5 using AF5795 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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. See Certificate of Analysis for details.
*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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Entrez Gene IDs

4907 (Human); 23959 (Mouse); 58813 (Rat); 102114976 (Cynomolgus Monkey)

Gene Symbol

NT5E

UniProt

AAH65937

Additional 5'-Nucleotidase/CD73 Products

Product Documents for Human 5'‑Nucleotidase/CD73 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human 5'‑Nucleotidase/CD73 Antibody

For research use only

Citations for Human 5'‑Nucleotidase/CD73 Antibody

Customer Reviews for Human 5'‑Nucleotidase/CD73 Antibody (1)

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  • Human 5'-Nucleotidase/CD73 Antibody
    Name: Anonymous
    Application: immunofluorescence - paraffin
    Sample Tested: human tonsil
    Species: Human
    Verified Customer | Posted 05/02/2019
    pH 9 heat-induced antigen retrieval
    Human 5'‑Nucleotidase/CD73 Antibody AF5795

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Protocols

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