Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag, CF

Biotinylated
Catalog # Availability Size / Price Qty
AVI5795-025
Biotinylated Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag Protein Binding Activity
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Citations (1)
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Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA.
When Human 5'-Nucleotidase/CD73 Antibody (Catalog # MAB5795) is coated at 1 μg/mL, 100 μL/well, Biotinylated Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag protein (Catalog # AVI5795) binds with an ED50 of 6-36 ng/mL.
Measured by its ability to hydrolyze the 5’-phosphate group from the substrate adenosine-5’-monophosphate (AMP). The orthophosphate product is measured by a Malachite Green Phosphate Detection Kit (Catalog # DY996). The specific activity is >15,000 pmol/min/μg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human 5'-Nucleotidase/CD73 protein
Human 5'-Nucleotidase/CD73
(Trp27 - Lys547)
Accession # AAH65937.1
Avi-tag6-His tag
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Trp27
Predicted Molecular Mass
61 kDa
SDS-PAGE
60-66 kDa, under reducing conditions.

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AVI5795

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI5795

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2 and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Scientific Data

Binding Activity View Larger

When Human 5'-Nucleotidase/CD73 Antibody (MAB5795) is coated at 1 μg/mL, 100 μL/well, Biotinylated Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag protein (Catalog # AVI5795) binds with an ED50 of 6-36 ng/mL.

SDS-PAGE View Larger

1 μg/lane of Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag Protein (Catalog # AVI5795) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 60-66 kDa.

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Background: 5'-Nucleotidase/CD73

CD73, known as ecto-5'-Nucleotidase, converts extracellular nucleoside‑5'‑monophosphates to nucleosides, with AMP as its preferred substrate (1). CD73 is a zinc-dependent, 70 kDa homodimeric enzyme bound to the cell membrane through a glycosyl phophatidylinositol (GPI) anchor. It is composed of an N-terminal domain containing metal binding sites linked via small hinge region to a C-terminal domain containing the substrate binding site and dimerization interface (2). It is expressed by most cell types (3) and is widely expressed in tumor cell lines as well as upregulated in cancerous tissues (4, 5). CD73 is a key enzyme responsible for a rate‑limiting step in the generation of extracellular adenosine. Adenosine is a molecule that signals through activation of purinergic receptors and results in an immunosuppressive role in the tumor microenvironment (4, 6). CD73 has been implicated in many pathological processes including immunomodulation and inflammation (7, 8) tumor growth and metastasis (9-13) making CD73 a potential drug target in cancer. Targeting CD73 inhibition has resulted in numerous reports of favorable antitumor effects (4, 5, 12). Consequently, therapeutic approaches have been tested using knockdown, gene silencing and anti-CD73 therapies (11, 14) as well as small molecule inhibitors (14, 15).

References
  1. Zimmermann, H. et al. (2012) Purinergic Signal. 8:437.
  2. Knapp, K. et al. (2012) Structure. 20:2161.
  3. Resta, R. et al. (1998) Immunol. Rev. 161:95. 
  4. Jin, D. et al. (2010) Cancer Res. 70:2245.
  5. Zhang, B. (2010) Cancer Res. 70:6407.
  6. Picher, M. et al. (2003) J. Biol. Chem. 278:13468. 
  7. Antonioli, L. et al. (2012) Curr. Drug Targets 13:842.
  8. Eltzschig, H. K. et al. (2012) N. Engl. J. Med. 367:2322.
  9. Bavaresco, L. et al. (2008) Mol. Cell Biochem. 319:61.
  10. Yegutkin, G. G. et al. (2011) Eur. J. Immunol. 41:1231.
  11. Stagg, J. et al. (2012) Cancer Res. 72:2190.
  12. Ghalamfarsa, G. et al. (2019) Expert Opin. Ther. Targets. 23:127.
  13. Gao, Z. W. et al. (2014) Biomed. Res. Int. [PMID 25126561].
  14. Young, A. et al. (2014) Cancer Discov. 4:879.
  15. McManus, J. et al. (2018) SLAS Discov. 23:264.
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

Citation for Recombinant Human 5'-Nucleotidase/CD73 His-tag Avi-tag, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Detection of CD39 and a Highly Glycosylated Isoform of Soluble CD73 in the Plasma of Patients with Cervical Cancer: Correlation with Disease Progression
    Authors: R Muñóz-Godí, M de Lourdes, B Weiss-Stei, JJ Montesinos, A Del Carmen, R García-Roc, J Hernández-, C Azucena Do, LR Ávila-Ibar, DB Torres-Pin, G Molina-Cas, R Chacón-Sal, L Vallejo-Ca, SM Pérez-Tapi, A Monroy-Gar
    Mediators of Inflammation, 2020;2020(0):1678780.
    Species: Human
    Sample Types: Reference Standard

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