Human 5'-Nucleotidase/CD73 Antibody

Name: Human 5'-Nucleotidase/CD73 Antibody
Brand: R&D Systems
SKU: AF5795

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF5795

Citations (2)

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Species Reactivity:	Human
Immunogen:	Chinese hamster ovary cell line CHO-derived recombinant human 5'‑Nucleotidase/CD73 Trp27-Lys547 Accession # AAH65937
Applications:	Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Simple Western, Western Blot
Label:	Unconjugated
Source:	Polyclonal Sheep IgG
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.

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Applications for Human 5'-Nucleotidase/CD73 Antibody

	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.

Scientific Data Examples for Human 5'-Nucleotidase/CD73 Antibody

	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.
	Immunocytochemistry 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. 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 Human 5'‑Nucleotidase/CD73 by Simple Western^TM. 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 Western^TM. 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.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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.

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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.

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

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:

Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

JV Korpershoe et al. (2021), 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, 22(16).
PMID: 34445320
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

EF Fang et al. (2019), NAD+ augmentation restores mitophagy and limits accelerated aging in Werner syndrome Nat Commun, 10(1):5284.
PMID: 31754102

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Human 5'-Nucleotidase/CD73 Antibody

R&D Systems, Inc. a Bio-Techne Brand

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Applications for Human 5'-Nucleotidase/CD73 Antibody

Scientific Data Examples for Human 5'-Nucleotidase/CD73 Antibody

Western Blot

Immunocytochemistry

Immunohistochemistry

Simple Western