Recombinant Human CD39L1/ENTPD2 Protein, CF Summary
Thr29-Asp460, with a C-terminal 6-His tag
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.
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.
|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.
- Assay Buffer: 25 mM Tris, 5 mM CaCl2, pH 7.5
- Recombinant Human CD39L1/ENTPD2 (rhCD39L1) (Catalog # 6087-EN)
- Substrate: ATP (Sigma, Catalog # A-7699), 10 mM stock 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
- Dilute rhCD39L1 to 0.04 µg/mL in Assay Buffer.
- 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).
- 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.
- Load 25 µL of 0.04 μg/mL rhCD39L1 and the standard curve into a plate. Include a Substrate Blank containing Assay Buffer.
- Dilute the Substrate to 100 µM in Assay Buffer.
- Add 25 µL of the 100 µM Substrate to all wells and mix well.
- Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 20 minutes.
- Add 10 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
- Add 10 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
- Read plate at 620 nm (absorbance) in endpoint mode.
- 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:
- rhCD39L1: 0.001 µg
- Substrate: 35.7 µM
- Phosphate Concentration: 2.5, 1.25, 0.625, 0.3125, 0.156, 0.078, and 0.039 nmol
CD39L1, also known as ENTPD2 and NTPDase2, is an ectonucleotidase belonging to the CD39 family. It is found on the surface of vascular adventitial cells and accessory vascular cells (1). CD39L1 is a Ca2+ and Mg2+ dependent enzyme that activates platelets by preferentially converting ATP to ADP (2). CD39L1 plays a role in regulating thrombosis and inflammation (3). It is considered to be a therapeutic target for thromboregulation and the treatment of vascular inflammation (2, 4).
Zimmermann, H. et al. 2000 Proceedings of the Second International Workshop on EctoATPases and Related Ectonucleotidases:18.
Robson, S.C. et al. 2001 Drug Dev. Res. 53:193.
Marcus, A.J. et al. 2005 Semin. Thromb. Hemost. 31:234.
Sevigny, J. et al. 2002 Blood 99:2801.
Citation for Recombinant Human CD39L1/ENTPD2 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
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.
Sample Types: Protein
Applications: ELISA Capture
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Phosphatase Activity Assays
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