Recombinant Human ENPP-1 Protein, CF
Recombinant Human ENPP-1 Protein, CF Summary
Product Specifications
| His | ENPP-2 Asp49-Trp144 Accession # Q13822 |
ENPP-1 Val191-Leu591 Accession # P22413 |
ENPP-2 Asn532-Ile863 Accession # Q13822 |
| N-terminus | C-terminus | ||
Analysis
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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.
6136-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 Tris, 250 mM NaCl, pH 9.5
- Recombinant Human ENPP-1 (rhENPP-1) (Catalog # 6136-EN)
- Substrate: Thymidine 5’-monophosphate p-nitrophenyl ester (Sigma, Catalog # T4510), 100 mM stock in deionized water
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhENPP-1 to 1 ng/µL in Assay Buffer.
- Dilute Substrate to 10 mM in Assay Buffer.
- Load into a plate 50 µL of 1 ng/µL rhENPP-1, and start the reaction by adding 50 µL of 10 mM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 10 mM Substrate.
- Read at 405 nm (absorbance) in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 4-Nitrophenol (Sigma-Aldrich, Catalog # 241326).
- rhENPP-1: 0.05 µg
- Substrate: 5 mM
Reconstitution Calculator
Background: ENPP-1
Ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP-1) is a transmembrane glycoprotein that hydrolyzes nucleotides and nucleotide derivatives with the formation of nucleotide-5’-monophosphates. It is inserted into the plasma membrane by an N‑terminal transmembrane domain. Human ENPP-1 has a small N-terminal cytoplasmic domain and a large C-terminal region containing two somatomedin B-like domains, a catalytic domain and a nuclease-like domain in the extracellular space (1). Defects in the ENPP-1 gene cause arterial calcification and bone mineralization abnormalities (2). ENPP-1 polymorphism or overexpression is also associated with obesity, type II diabetes and insulin resistance, which makes modulation of ENPP-1 activity one of the targets to treat insulin resistance and related diseases (1). The recombinant hybrid enzyme recombinant human ENPP-1 consists of N-terminal somatomedin B-like domains of ENPP-2, the central catalytic phosphodiesterase domain of ENPP-1 and the C‑terminal nuclease-like domain of ENPP-2. It has been reported that this hybrid construct generates an enzyme that has higher levels of ENPP-1 activity than the wild type ENPP-1 and is specific for the ENPP-1 substrate (3).
- Goldfine, I.D. et al. (2010) Endocrine Reviews. 29:62.
- Hessle, L. et al. (2002) Proc. Natl. Acad. Sci. 99:9445.
- Anisoara, C. et al. (2004) Biochem. J. 381:71.
Citation for Recombinant Human ENPP-1 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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Non-invasive imaging of tumors by monitoring autotaxin activity using an enzyme-activated near-infrared fluorogenic substrate.
Authors: Madan, Damian, Ferguson, Colin G, Lee, Won Yong, Prestwich, Glenn D, Testa, Charles
PLoS ONE, 2013;8(11):e79065.
Species: N/A
Sample Types: Complex Sample Type
Applications: Bioassay
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Reason for Rating: Worked beautifully in dose-response assay.
