Human 5'-Nucleotidase/CD73 Antibody

Catalog # Availability Size / Price Qty
AF5795
AF5795-SP
Human 5'-Nucleotidase/CD73 Antibody in Western Blot
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Human 5'-Nucleotidase/CD73 Antibody Summary

Species Reactivity
Human
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.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human 5'‑Nucleotidase/CD73
Trp27-Lys547
Accession # AAH65937
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
1-15 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
See below

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.

Data Examples

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

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.

Immunocytochemistry 5'‑Nucleotidase/CD73  in A431 Human Cell Line. View Larger

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.

Immunohistochemistry 5'‑Nucleotidase/CD73  in Human Cervical Cancer Tissue. View Larger

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.

Simple Western Detection of Human5'‑Nucleotidase/CD73 by Simple WesternTM. View Larger

Detection of Human
5'‑Nucleotidase/CD73 by Simple WesternTM.
Simple Western lane view shows lysates of U266 human myeloma 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) 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.

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

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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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.

Entrez Gene IDs
4907 (Human); 23959 (Mouse); 58813 (Rat); 102114976 (Cynomolgus Monkey)
Alternate Names
5' nucleotidase (CD73); 5'-NT; 5-NT; 5'-Nucleotidase; 5'-nucleotidase, ecto (CD73); CD73 antigen; CD73; E5NT; EC 3.1.3.5; ecto-5'-nucleotidase; eN; eNT; NT; NT55'-nucleotidase; NT5E; NTE; Purine 5-Prime-Nucleotidase

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FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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