Recombinant Human CD39/ENTPD1 Protein, CF
Recombinant Human CD39/ENTPD1 Protein, CF Summary
Thr38-Val478, with a C-terminal 6-His tag
|Formulation||Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2 and Glycerol.|
|Shipping||The product is shipped with dry ice or equivalent. 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 CD39/ENTPD1 (rhCD39) (Catalog # 4397-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 rhCD39 to 0.04 µg/mL in Assay Buffer.
- 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.
- Transfer to a 96 well clear plate (in duplicate) 25 µL of diluted rhCD39 at 0.04 µg/mL, standard curve, and blanks (Assay Buffer).
- Dilute Substrate to 100 µM in Assay Buffer.
- Add 25 µL of the diluted 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/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:
- rhCD39: 0.001 µg
- Substrate: 35.7 µM
Ectonucleoside triphosphate diphosphohydrolase-1 (NTPDase-1) is an integral membrane protein with an extracellular active site. Recombinant human NTPDase-1 was expressed as a protein lacking its N- and C-terminal transmembrane domains, resulting in the secretion of the soluble recombinant human NTPDase-1 ectodomain. NTPDase-1 was originally described as CD39, a B lymphocyte cell surface marker (2), but it is also present on the surface of natural killer cells, T cells, and some endothelial cells (3). 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 (4). NTPDase-1 is involved in the processes of thromboregulation and vascular inflammation (5). The administration of soluble NTPDase-1 may have therapeutic applications for the treatment of some vascular and transplantation-associated diseases (6).
- Maliszewski, C.R. et al. (1994) J. Immunol. 153:3574.
- 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.
Citations for Recombinant Human 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.
Citations: Showing 1 - 2
Filter your results:
ADPase CD39 Fused to Glycoprotein VI-Fc Boosts Local Antithrombotic Effects at Vascular Lesions
Authors: H Degen, O Borst, M Ziegler, AK Mojica Mun, J Jamasbi, B Walker, S Göbel, J Fassbender, K Adler, R Brandl, G Münch, R Lorenz, W Siess, M Gawaz, M Ungerer
J Am Heart Assoc, 2017;6(8):.
The ubiquitin ligase ASB4 promotes trophoblast differentiation through the degradation of ID2.
Authors: Townley-Tilson, W H Davi, Wu, Yaxu, Ferguson, James E, Patterson, Cam
PLoS ONE, 2014;9(2):e89451.
Sample Types: Whole Cells
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
Phosphatase Activity Assays
Protein Purification Kits
Reviews for Recombinant Human CD39/ENTPD1 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human CD39/ENTPD1 Protein, CF and earn rewards!
Have you used Recombinant Human CD39/ENTPD1 Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image