Calcium Activated Nucleotidase 1 (CANT1), is a Ca2+-dependent nucleoside diphosphatase which can hydrolyze a variety of nucleoside di- and triphosphates. CANT1 is also known as SCAN-1 and SHAPY. Its preferred substrates are UDP and GDP, and it is not active against nucleoside monophosphates (1). The CANT1 gene encodes a Type II membrane protein that is anchored in the endoplasmic reticulum (ER) membrane with its catalytic domain in the ER lumen (2). A secreted, soluble form of the enzyme also exists (1). CANT1 is expressed in many tissues, including fibroblasts and chondrocytes. Mutations of CANT1 are a cause of Desbuquois dysplasia, a condition characterized by deformation of bones and joints (2, 3). Recombinant human CANT1 was expressed as a secreted, soluble protein.
Recombinant Human CANT1 Protein, CF
R&D Systems | Catalog # 6720-NT
Key Product Details
- R&D Systems NS0-derived Recombinant Human CANT1 Protein (6720-NT)
- 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
Source
Accession Number
Applications
Product Specifications
Source
Gly80-Ile401, with an N-terminal 6-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
Measured by its ability to hydrolyze the beta 5'-phosphate group from the substrate uridine-5'-diphosphate (UDP). The orthophosphate product is measured by a Malachite Green Phosphate Detection Kit (Catalog # DY996).
The specific activity is >85,000 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
6720-NT
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl 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.
|
Background: Calcium Activated Nucleotidase 1/CANT1
References
- Smith, T. et al. (2002) Arch. Biochim. Biophys. 406:105.
- Failer, B.U. et al. (2002) J. Biol. Chem. 277:36978.
- Huber, C. et al. (2009) Am. J. Hum. Genet. 85:706.
- Faden, M. et al. (2010) Am. J. Med. Genet. A. 152A:1157.
Long Name
Alternate Names
Gene Symbol
UniProt
Additional Calcium Activated Nucleotidase 1/CANT1 Products
Product Documents for Recombinant Human CANT1 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human CANT1 Protein, CF
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human CANT1 Protein, CF (6720-NT):
- Assay Buffer: 50 mM MES, 5 mM CaCl2, pH 6.5
- Recombinant Human Calcium Activated Nucleotidase 1/CANT1 (rhCANT1) (Catalog # 6720-NT)
- Substrate: Uridine 5’-diphosphate sodium salt (UDP) (Sigma, Catalog # U4125)
- 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 1 M Phosphate Standard by adding 10 µL of the 1 M Phosphate Standard to 990 µL of deionized water 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.
- Prepare standard curve by performing seven 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.
- Dilute rhCANT1 to 0.005 µg/mL in Assay Buffer.
- Dilute Substrate to 100 µM in Assay Buffer.
- Load 50 µL of each dilution of the standard curve into a plate. Include a Curve Blank containing 50 μL of Assay Buffer.
- Load 25 µL of the 0.005 µg/mL rhCANT1 into the plate. Include a Substrate Blank containing 25 µL of Assay Buffer.
- Add 25 µL of 100 µM Substrate to the wells, excluding the standard curve and Curve Blank.
- 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 Reaction:
- rhCANT1: 0.000125 µg
- UDP: 50 µM