Recombinant Human CD39L2/ENTPD6 Protein, CF

Catalog # Availability Size / Price Qty
4399-EN-010
Product Details
Citations (1)
FAQs
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Recombinant Human CD39L2/ENTPD6 Protein, CF Summary

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 guanosine-5’-diphosphate (GDP). The orthophosphate product is measured by a Malachite Green Phosphate Detection Kit (Catalog # DY996). The specific activity is >30,000 pmol/min/µg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human CD39L2/ENTPD6 protein
Lys61-Ser484, with an N-terminal 6-His tag
Accession # O75354
Accession #
N-terminal Sequence
Analysis
His
Predicted Molecular Mass
47 kDa
SDS-PAGE
50-55 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.

4399-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.

Assay Procedure

Materials
  • Assay Buffer: 25 mM Tris, 5 mM CaCl2, pH 7.5
  • Recombinant Human CD39L2/ENTPD6 (rhCD39L2) (Catalog # 4399-EN)
  • Substrate: GDP (Sigma, Catalog # G-7127), 100mM stock in deionized H2O
  • 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 rhCD39L2 to 0.02 µg/mL in Assay Buffer.
  2. Prepare a standard curve from 1 M Phosphate Standard supplied in the Malachite Green Phosphate Detection Kit:
    1. Add 10 µL of the 1 M Phosphate Standard to 990 µL of Assay Buffer for a 10 mM stock.
    2. Add 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.
  3. Load 25 µL of 0.02 μg/mL rhCD39L2 and the standard curve into a plate. Include a Substrate Blank containing Assay Buffer.
  4. Dilute the Substrate to 100 µM in Assay Buffer.
  5. Add 25 µL of the 100 µM Substrate to all wells and mix well.
  6. Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 20 minutes.
  7. Add 10 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
  8. Add 10 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  9. Read plate at 620 nm (absorbance) in endpoint mode.
  10. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Phosphate released* (nmol) x (1000 pmol/1 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:
  • rhCD39L2:  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: CD39L2/ENTPD6

Ectonucleoside triphosphate diphosphohydrolase-6 (NTPDase-6) is a secreted nucleoside phosphohydrolase of the CD39 family of enzymes (1). hNTPDase-6 displays a preference for the nucleoside-5’-diphosphates GDP and IDP over CDP and UDP (2). Nucleoside-5’-triphosphates are also hydrolyzed, but at much lower rates. hNTPDase-6 has a broad tissue distribution, its mRNA has been detected in all human tissues tested (1).

References
  1. Chadwick, B.P. and A.M. Frischauf (1998) Genomics. 50:357.
  2. Hicks-Berger, C.A. et al. (2000) J. Biol. Chem. 275:34041.
Long Name
Ecto-Nucleoside Triphosphate Diphosphohydrolase 6
Entrez Gene IDs
955 (Human); 12497 (Mouse); 85260 (Rat)
Alternate Names
CD39 antigen-like 2; CD39L2; CD39L2NTPDase-6; CD39-like 2; dJ738P15.3; DKFZp781G2277; DKFZp781K21102; EC 3.6.1.6; ectonucleoside triphosphate diphosphohydrolase 6 (putative function); ectonucleoside triphosphate diphosphohydrolase 6 (putative); ectonucleoside triphosphate diphosphohydrolase 6; ENTPD6; FLJ36711; IL-6SAG; IL6ST2; interleukin 6 signal transducer-2; NTPD3; NTPDase 6

Citation for Recombinant Human CD39L2/ENTPD6 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.

1 Citation: Showing 1 - 1

  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

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