Recombinant Mouse CD39/ENTPD1 Protein, CF Summary
- R&D Systems NS0-derived Recombinant Mouse CD39/ENTPD1 Protein (4398-EN)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Product Specifications
Thr38-Ile478, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
4398-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.
|
Assay Procedure
- Assay Buffer: 25 mM Tris, 5 mM CaCl2, pH 7.5
- Recombinant Mouse CD39/ENTPD1 (rmCD39) (Catalog # 4398-EN)
- Substrate: Adenosine triphosphate (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
- Prepare a standard curve from the 1 M Phosphate Standard supplied in the malachite green phosphate detection kit. First, add 10 µL of the 1 M Phosphate Standard to 990 µL of Assay Buffer for a 10 mM stock. Then, 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). Finally, perform six additional two-fold serial dilutions of the 100 µM phosphate stock. The standard curve has a range of 0.039 to 2.5 nmol per well.
- Dilute rmCD39 to 0.02 µg/mL in Assay Buffer.
- Transfer to a plate (in duplicate) 25 µL of standard curve, diluted rmCD39 at 0.02 µg/mL, and blanks (Assay Buffer).
- Dilute Substrate to 100 µM in Assay Buffer.
- Add 25 µL of the Substrate to all wells. Mix well.
- Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 30 minutes.
- After incubation, add 10 µL of the Malachite Green Reagent A to each sample, standard, and blank. Mix and incubate for 10 minutes at room temperature.
- Add 10 µL of the Malachite Green Reagent B to each sample, standard, and blank. 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 Reaction:- rmCD39: 0.0005 µg
- Substrate: 50 µM
Reconstitution Calculator
Background: CD39/ENTPD1
Ectonucleoside triphosphate diphosphohydrolase-1 (NTPDase-1) is an integral membrane protein with an extracellular active site. Recombinant mouse NTPDase-1 was expressed as a protein lacking its N- and C-terminal transmembrane domains, resulting in the secretion of the soluble ectodomain. NTPDase-1 was originally described as CD39, a B lymphocyte cell surface marker (1), but it is also present on the surface of natural killer cells, T cells, and some endothelial cells (2). NTPDase-1 hydrolyzes the beta - and gamma phosphate residues of nucleotides, preferring ATP as the substrate. Through its hydrolysis of extracellular nucleotides, NTPDase-1 plays a role in the regulation of purinergic signaling (3). NTPDase-1 is involved in the processes of thromboregulation and vascular inflammation (4). The administration of soluble NTPDase-1 may have therapeutic applications for the treatment of some vascular and transplantation-associated diseases (5).
- Rowe, M. et al. (1982) Int. J. Cancer 29:373.
- Kansas, G.S. et al. (1991) J. Immunol. 146:2235.
- Kishore, B.K. et al. (2005) Am. J. Physiol. Renal Physiol. 288:F1032.
- Marcus, A.J. et al. (2005) Semin. Thromb. Hemost. 31:234.
- Robson, S.C. et al. (2005) Semin. Thromb. Hemost. 31:217.
Citation for Recombinant Mouse CD39/ENTPD1 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
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Synergistic effect of tumor chemo-immunotherapy induced by leukocyte-hitchhiking thermal-sensitive micelles
Authors: J Qi, F Jin, Y You, Y Du, D Liu, X Xu, J Wang, L Zhu, M Chen, G Shu, L Wu, J Ji, Y Du
Nature Communications, 2021-08-06;12(1):4755.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo
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