Recombinant Human Alkaline Phosphatase/ALPL Protein, CF

Catalog # Availability Size / Price Qty
2909-AP-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (3)
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Recombinant Human Alkaline Phosphatase/ALPL Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to cleave a fluorogenic substrate, 4-Methylumbelliferyl phosphate (4-MUP). The specific activity is >30,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Alkaline Phosphatase/ALPL protein
Leu18-Ser502, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Leu18
Predicted Molecular Mass
54 kDa
SDS-PAGE
65-75 kDa, reducing conditions

Product Datasheets

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2909-AP

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

2909-AP

Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, MgCl2 and ZnCl2.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 1 mM MgCl2, pH 9.0
  • Recombinant Human Alkaline Phosphatase/ALPL/TNAP (rhTNAP) (Catalog # 2909-AP)
  • Substrate: 4-Methumbelliferly phosphate (4-MUP) (Calbiochem, Catalog # 474431), 10 mM in deionized water
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhTNAP to 0.01 ng/µL in Assay Buffer.
  2. Dilute Substrate 50 µM in Assay Buffer.
  3. In a plate load 50 µL of 0.01 ng/µL rhTNAP to wells. Include a Substrate Blank of 50 µL of Assay Buffer.
  4. Start the reaction by adding 50 µL of 50 µM Substrate to wells.
  5. Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively in kinetic mode for 5 minutes.
  6. 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).

Per Well:
  • rhTNAP: 0.0005 µg
  • Substrate: 25 µM
Reconstitution Calculator

Reconstitution Calculator

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Background: Alkaline Phosphatase/ALPL

Four distinct genes encode alkaline phosphatases (APs) in humans (1). The ALPL gene encodes the liver/bone/kidney isozyme, also known as the tissue‑nonspecific AP (TNAP). In comparison, ALPI, ALPP and ALPPL2 encode intestinal, placental and placental-like or germ cell APs, respectively. The serum levels of human APs are useful tumor markers (2). There are many mutations in the ALPL gene, leading 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 503 to 524) is not present in the mature form.

References
  1. Le Du, M.-H. and J.L. Millan (2002) J. Biol. Chem. 277:49808.
  2. Millan, J.L. and W.H. Fishman (1995) Crit. Rev. Clin. Lab. Sci. 32:1.
  3. Di Mauro, S. et al. (2002) J. Bone Miner. Res. 17:1383.
Long Name
Alkaline Phosphatase Liver
Entrez Gene IDs
249 (Human); 11647 (Mouse)
Alternate Names
Akp2; Alkaline phosphatase liver/bone/kidney isozyme; alkaline phosphatase, liver/bone/kidney; alkaline phosphatase, tissue-nonspecific isozyme; alkaline phosphomonoesterase; ALPL; APTNAP; AP-TNAP; EC 3.1.3.1; FLJ40094; FLJ93059; glycerophosphatase; HOPS; liver/bone/kidney-type alkaline phosphatase; MGC161443; tissue-nonspecific ALP; TNAP; TNSALP; TNSALPMGC167935

Citations for Recombinant Human Alkaline Phosphatase/ALPL 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.

3 Citations: Showing 1 - 3
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  1. Ocular Hypotensive Properties and Biochemical Profile of QLS-101, a Novel ATP-Sensitive Potassium (KATP) Channel Opening Prodrug
    Authors: CL Pervan-Ste, U Roy Chowdh, HK Sookdeo, RA Casale, PI Dosa, TM Htoo, MP Fautsch, BM Wirostko
    Investigative Ophthalmology & Visual Science, 2022;63(4):26.
    Species: Human
    Sample Types: Small Molecule
    Applications: Bioassay
  2. Mitochondrial TNAP controls thermogenesis by hydrolysis of phosphocreatine
    Authors: Y Sun, JF Rahbani, MP Jedrychows, CL Riley, S Vidoni, D Bogoslavsk, B Hu, PA Dumesic, X Zeng, AB Wang, NH Knudsen, CR Kim, A Marasciull, JL Millán, ET Chouchani, L Kazak, BM Spiegelman
    Nature, 2021;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Bioassay
  3. Human alkaline phosphatase dephosphorylates microbial products and is elevated in preterm neonates with a history of late-onset sepsis
    Authors: M Pettengill, JD Matute, M Tresenrite, J Hibbert, D Burgner, P Richmond, J Luis Millá, A Ozonoff, T Strunk, A Currie, O Levy
    PLoS ONE, 2017;12(4):e0175936.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay

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