Recombinant Human CD39L3/ENTPD3 Protein, CF

Catalog # Availability Size / Price Qty
4400-EN-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human CD39L3/ENTPD3 Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze the 5’-phosphate groups from the substrate adenosine-5’-triphosphate (ATP). The orthophosphate product is measured by a Malachite Green Phosphate Detection Kit (Catalog # DY996). The specific activity is >70,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human CD39L3/ENTPD3 protein
Gln44-Pro485, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
No results obtained: Gln44 predicted
Predicted Molecular Mass
50 kDa
SDS-PAGE
65-80 kDa, reducing conditions

Product Datasheets

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.

4400-EN

Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl 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.

Assay Procedure

Materials
  • Assay Buffer: 25 mM Tris, 5 mM CaCl2, pH 7.5
  • Recombinant Human CD39L3/ENTPD3 (rhCD39L3) (Catalog # 4400-EN)
  • Substrate: ATP (Sigma, Catalog # A-7699), 10 mM in deionized water
  • Malachite Green Phosphate Detection Kit (Catalog # DY996)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhCD39L3 to 0.02 µg/mL in Assay Buffer.
  2. Prepare a standard curve from 1 M Phosphate Standard by adding 10 µL of the 1 M Phosphate Standard to 990 µL of Assay Buffer for a 10 mM stock. Continue by adding 10 µL of the 10 mM Phosphate stock to 990 µL of Assay Buffer for a 100 µM stock. (This is the first dilution to use as a standard).
  3. Perform six additional one-half serial dilutions of the 100 µM Phosphate stock in Assay Buffer. The standard curve has a range of 0.039 to 2.5 nmol per well.
  4. Load 25 µL of 0.02 µg/mL rhCD39L3 and the standard curve into a plate. Include a Substrate Blank containing Assay Buffer.
  5. Dilute the Substrate to 100 µM in Assay Buffer.
  6. Add 25 µL of the 100 µM Substrate to all wells and mix well.
  7. Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 30 minutes.
  8. Add 10 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
  9. Add 10 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  10. Read plate at 620 nm (absorbance) in endpoint mode.
  11. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Phosphate released* (nmol) x (1000 pmol/nmol)
Incubation time (min) x amount of enzyme (µg)

     *Derived from the phosphate standard curve using linear or 4-parameter fitting and adjusted for Substrate Blank.

Per Well:
  • rhCD39L3: 0.0005 µg
  • Substrate: 35.7 µM
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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Background: CD39L3/ENTPD3

Ectonucleoside triphosphate diphosphohydrolase-3 (NTPDase-3), encoded by the ENTPD3 gene and also known as CD39L3, is an integral membrane protein with an extracellular active site (1). Recombinant human NTPDase-3 was expressed as a protein lacking its N- and C-terminal transmembrane domains, resulting in the secretion of the soluble ectodomain. NTPDase-3 hydrolyzes the beta - and gamma -phosphate residues of nucleotides, preferring ATP, ADP, UTP, and UDP as substrates (1). Through its hydrolysis of extracellular nucleotides, NTPDase-3 plays a role in the regulation of purinergic signaling (2). The enzyme is expressed at its highest levels in brain, pancreas, spleen and prostate tissues (3).

References
  1. Lavoie, E.G. et al. (2004) Biochem. Pharmacol. 67:1917.
  2. Crawford, P.A. et al. (2007) Arch. Biochem. Biophys. 457:7.
  3. Chadwick, B.P. and A.M. Frischauf (1998) Genomics 50:357.
Long Name
Ecto-Nucleoside Triphosphate Diphosphohydrolase 3
Entrez Gene IDs
956 (Human); 215446 (Mouse); 316077 (Rat)
Alternate Names
ATPDase; CD39 antigen-like 3; CD39L3; CD39L3EC 3.6.1.5; CD39-like 3; EC 3.6.1; Ecto-apyrase 3; Ecto-ATP diphosphohydrolase 3; Ecto-ATPase 3; Ecto-ATPDase 3; ectonucleoside triphosphate diphosphohydrolase 3; ENTPD3; FLJ93839; HB6; HB6NTPDase 3; NTPDase-3

Citations for Recombinant Human CD39L3/ENTPD3 Protein, 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.

3 Citations: Showing 1 - 3
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  1. Blocking Antibodies Targeting the CD39/CD73 Immunosuppressive Pathway Unleash Immune Responses in Combination Cancer Therapies
    Authors: I Perrot, HA Michaud, M Giraudon-P, S Augier, A Docquier, L Gros, R Courtois, C Déjou, D Jecko, O Becquart, H Rispaud-Bl, L Gauthier, B Rossi, S Chanteux, N Gourdin, B Amigues, A Roussel, A Bensussan, JF Eliaou, J Bastid, F Romagné, Y Morel, E Narni-Manc, E Vivier, C Paturel, N Bonnefoy
    Cell Rep, 2019;27(8):2411-2425.e9.
    Species: Human
    Sample Types: Protein
    Applications: ELISA Capture
  2. Phostine PST3.1a Targets MGAT5 and Inhibits Glioblastoma Initiating Cell Invasiveness and Proliferation
    Authors: Z Hassani, A Saleh, S Turpault, S Khiati, W Morelle, J Vignon, JP Hugnot, E Uro-Coste, P Legrand, M Delaforge, S Loiseau, L Clarion, M Lecouvey, JN Volle, D Virieux, JL Pirat, H Duffau, N Bakalara
    Mol. Cancer Res., 2017;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  3. The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor DELLA
    Authors: R Zentella, N Sui, B Barnhill, WP Hsieh, J Hu, J Shabanowit, M Boyce, NE Olszewski, P Zhou, DF Hunt, TP Sun
    Nat. Chem. Biol, 2017;0(0):.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay

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