Source	Chinese Hamster Ovary cell line, CHO-derived cynomolgus monkey 5'-Nucleotidase/CD73 protein Trp27-Lys547 with a C-terminal 6-His tag
Purity	>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Trp27
Predicted Molecular Mass	59 kDa
SDS-PAGE	57-75 kDa
Activity	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 >6500 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation and Storage

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.

10173-EN

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.

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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).

Item	Value
References	Zimmermann, H. et al. (2012) Purinergic Signal. 8:437. Knapp, K. et al. (2012) Structure. 20:2161. Resta, R. et al. (1998) Immunol. Rev. 161:95. Jin, D. et al. (2010) Cancer Res. 70:2245. Zhang, B. (2010) Cancer Res. 70:6407. Pilcher, M. et al. (2003) J. Biol. Chem. 278:13468. Antonioli, L. et al. (2012) Curr. Drug Targets 13:842. Eltzchig, H. K. et al. (2012) N. Engl. J. Med. 367:2322. Bavaresco, L. et al. (2008) Mol. Cell Biochem. 319:61. Yegutkin, G. G. et al. (2011) Eur. J. Immunol. 41:1231. Stagg, J. et al. (2012) Cancer Res. 72:2190. Ghalamfarsa, G. et al. (2019) Expert Opin. Ther. Targets. 23:127. Gao, Z. W. et al. (2014) Biomed. Res. Int. [PMID 25126561]. Young, A. et al. (2014) Cancer Discov. 4:879. 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'-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

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Recombinant Cynomolgus 5'-Nucleotidase/CD73 His Protein, CF

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Background: 5'-Nucleotidase/CD73

Citations for Recombinant Cynomolgus 5'-Nucleotidase/CD73 His Protein, CF

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