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Recombinant Human ENPP-5 Protein, CF (20 ug)
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咨询Recombinant Human ENPP-5 Protein, CF Summary
- R&D Systems HEK293-derived Recombinant Human ENPP-5 Protein (8655-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
Pro25-Ser430, with a C-terminal 10-His tag
Analysis
Product Datasheets
8655-EN
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.
8655-EN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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: 50 mM Sodium Acetate, 150 mM NaCl, pH 5.5
- Recombinant Human ENPP-5 (rhENPP-5) (Catalog # 8655-EN)
- Substrate: O-(4-Nitrophenylphosphoryl) choline (Sigma, Catalog # N5879), 500 mM stock in deionized water
- NaOH, 0.2 M in deionized water
- 96-well Clear Plate (Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhENPP-5 to 5 µg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load 50 µL of 5 µg/mL rhENPP-5 in a clear strip well plate, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL 2 mM Substrate.
- Incubate sealed plate at room temperature for 30 minutes.
- Stop reactions by adding 100 µL of 0.2 M NaOH to each well used.
- Read at 410 nm (absorbance) in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Abs* (OD) x Conversion Factor** (pmol/OD) |
| Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard p-Nitrophenol (Sigma, Catalog # 241326).
- rhENPP-5: 0.25 µg
- Substrate: 1 mM
Reconstitution Calculator
Background: ENPP-5
Ectonucleotide pyrophosphatase/phosphodiesterase 5 (ENPP-5 or NPP5) belongs to a group of ecto-enzymes which regulate the availability of extracellular nucleotides (1). This enzyme family forms a subgroup of a larger family that also includes arylsulfatases, phosphopentomutases, 2,3-bisphosphoglycerate-independent phosphoglycerate mutases (iPGM), and alkaline phosphatases (2). Mature human ENPP-5 is an approximately 50 kDa protein that consists of a 407 amino acid (aa) extracellular region with one phosphodiesterase/nucleotide pyrophosphatase domain, a 21 aa transmembrane segment, and a 25 aa cytoplasmic tail (3, 4). Within the ectodomain, human ENPP-5 shares 85% aa sequence identity with mouse and rat ENPP-5. R&D Systems in-house testing indicates that recombinant human ENPP-5 exhibits phosphodiesterase activity with the substrate O-(4-Nitrophenylphosphoryl) choline.
- Zimmermann, H. et al. (2012) Purinergic Signal. 8:437.
- Gijsbers, R. et al. (2001) J. Biol. Chem. 276:1361.
- Ohe, Y. et al. (2003) Biochem. Biophys. Res. Commun. 308:719.
Background: ENPP-5
Ectonucleotide pyrophosphatase/phosphodiesterase 5 (ENPP-5 or NPP5) belongs to a group of ecto-enzymes which regulate the availability of extracellular nucleotides (1). This enzyme family forms a subgroup of a larger family that also includes arylsulfatases, phosphopentomutases, 2,3-bisphosphoglycerate-independent phosphoglycerate mutases (iPGM), and alkaline phosphatases (2). Mature human ENPP-5 is an approximately 50 kDa protein that consists of a 407 amino acid (aa) extracellular region with one phosphodiesterase/nucleotide pyrophosphatase domain, a 21 aa transmembrane segment, and a 25 aa cytoplasmic tail (3, 4). Within the ectodomain, human ENPP-5 shares 85% aa sequence identity with mouse and rat ENPP-5. R&D Systems in-house testing indicates that recombinant human ENPP-5 exhibits phosphodiesterase activity with the substrate O-(4-Nitrophenylphosphoryl) choline.
- Zimmermann, H. et al. (2012) Purinergic Signal. 8:437.
- Gijsbers, R. et al. (2001) J. Biol. Chem. 276:1361.
- Ohe, Y. et al. (2003) Biochem. Biophys. Res. Commun. 308:719.