Human 5'-Nucleotidase/CD73 Antibody
R&D Systems, part of Bio-Techne | Catalog # AF5795
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Dual RNAscope ISH-IHC Compatible, Immunocytochemistry, Immunohistochemistry, Simple Western, Western Blot
Cited:
Flow Cytometry, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
Product Specifications
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
Dual RNAscope ISH-IHC Compatible
5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer
Sample: Immersion fixed paraffin-embedded sections of human colon cancer
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:
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.
*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.
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
Gene Symbol
NT5E
UniProt
Additional 5'-Nucleotidase/CD73 Products
- All Products for 5'-Nucleotidase/CD73
- 5'-Nucleotidase/CD73 Activity Assays
- 5'-Nucleotidase/CD73 cDNA Clones
- 5'-Nucleotidase/CD73 ELISA Kits
- 5'-Nucleotidase/CD73 Lysates
- 5'-Nucleotidase/CD73 Primary Antibodies
- 5'-Nucleotidase/CD73 Proteins and Enzymes
- 5'-Nucleotidase/CD73 Small Molecules and Peptides
Product Documents for Human 5'-Nucleotidase/CD73 Antibody
Product Specific Notices for Human 5'-Nucleotidase/CD73 Antibody
For research use only
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