Several distinct genes encode alkaline phosphatases (APs) in mice with different tissue-specific expression patterns. The Alpl gene, also known as Akp2, encodes the liver/bone/kidney isozyme, also known as the tissue-nonspecific AP (TNAP) (1). The Alpl gene is a key regulator of bone mineralization in mice (2). A variety of mutations in the human ALPL gene leads to different forms of hypophosphatasia, characterized by poorly mineralized cartilage and bones (3). The native ALPL is a glycosylated homodimer attached to the membrane through a GPI-anchor. The C-terminal pro peptide (residues 504 to 524) is not present in the mature form.
Recombinant Mouse Alkaline Phosphatase/ALPL Protein, CF
R&D Systems | Catalog # 2910-AP
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Alkaline Phosphatase/ALPL Protein (2910-AP)
- 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
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Alkaline Phosphatase/ALPL protein
Phe18-Gly503, with a C-terminal 6-His tag
Phe18-Gly503, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Phe18
Predicted Molecular Mass
54 kDa
SDS-PAGE
77 kDa, reducing conditions
Activity
Measured by its ability to cleave a fluorogenic substrate, 4-Methylumbelliferyl phosphate (4-MUP).
The specific activity is >40,000 pmol/min/µg, as measured under the described conditions.
The specific activity is >40,000 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2910-AP
| 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.
|
Background: Alkaline Phosphatase/ALPL
References
- Terao, M. and B. Mintz (1987) Proc. Natl. Acad. Sci. USA 84:7051.
- Hessle, L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:9445.
- Di Mauro, S. et al. (2002) J. Bone Miner. Res. 17:1383.
Long Name
Alkaline Phosphatase Liver
Alternate Names
Akp2, AP-TNAP, HOPS, TNAP, TNSALP
Gene Symbol
ALPL
UniProt
Additional Alkaline Phosphatase/ALPL Products
Product Documents for Recombinant Mouse Alkaline Phosphatase/ALPL Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Alkaline Phosphatase/ALPL Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Alkaline Phosphatase/ALPL Protein, CF (2910-AP):
Materials
- Assay Buffer: 50 mM Tris, 1 mM MgCl2, pH 9.0
- Recombinant Mouse Alkaline Phosphatase/ALPL (rmALPL) (Catalog # 2910-AP)
- Substrate: 4-Methylumbelliferlyphosphate (4-MUP) (Sigma, Catalog # M8883)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmALPL to 0.02 ng/µL in Assay Buffer.
- Dilute Substrate to 50 µM in Assay Buffer.
- Load 50 µL of 0.02 ng/µL rmALPL into a plate, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
- Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively, in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 4-Methumbelliferone (Sigma, Catalog # M1381).
- rmALPL: 0.001 µg
- Substrate: 25 µM
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