Mouse Fetuin A, also known as alpha 2-Heremans-Schmid glycoprotein, is encoded by the AHSG gene. It has been also called "countertrypsin" because of its ability to inhibit trypsin (1). It is a major plasma protein and a member of the cystatin superfamily of protease inhibitors (2, 3). It is expressed by hepatocytes, the principal cell source, and by monocyte/macrophages (4). The major form of plasma Fetuin A corresponds to two disulfide bond-linked chains derived from the single chain (5). Fetuin-A has a number of functions. It is a negative acute-phase protein with normal circulating levels in adults (300-600 μg/mL), which fall significantly (30-50%) during injury and infection (5). It enhances entry of cationic inhibitors into macrophages (6). It inhibits both insulin receptor autophosphorylation and undesirable calcification (7, 8).
Recombinant Mouse Fetuin A/AHSG Protein, CF
R&D Systems | Catalog # 1563-PI
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Fetuin A/AHSG Protein (1563-PI)
- 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
Inhibition Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Fetuin A/AHSG protein
Ala19-Ile345, with a C-terminal 10-His tag
Ala19-Ile345, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Ala19
Predicted Molecular Mass
37 kDa
SDS-PAGE
58 kDa, reducing conditions
Activity
Measured by its ability to inhibit trypsin cleavage of a fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The IC50 value is <0.20 µM, as measured under the described conditions.
The IC50 value is <0.20 µM, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 1563-PI in the following applications:
Formulation, Preparation, and Storage
1563-PI
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 500 μg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 7.5. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Calculators
Background: Fetuin A/AHSG
References
- Yamamoto, K. and H. Sinohara (1993) J. Biol. Chem. 268:17750.
- Kellemann, J. et al. (1989) J. Biol. Chem. 264:14121.
- Dziegielewska, K.M. et al. (1990) J. Biol. Chem. 265:4354.
- Dziegielewska, K.M. et al. (1996) Histochem. Cell Biol. 106:319.
- Gejyo, F. and K. Schmid (1981) Biochim. Biophys. Acta 671:78.
- Wang, H. et al. (1998) Proc. Natl. Acad. Sci. USA 95:14429.
- Mathews, S.T. et al. (2000) Mol. Cell Endocrinol. 164:87.
- Schäfer, C. et al. (2003) J. Clin. Invest. 112:357.
Long Name
alpha-2-HS Glycoprotein
Alternate Names
AHSG, alpha-2-HS-glycoprotein
Gene Symbol
AHSG
UniProt
Additional Fetuin A/AHSG Products
Product Documents for Recombinant Mouse Fetuin A/AHSG 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 Mouse Fetuin A/AHSG Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse Fetuin A/AHSG Protein, CF
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Protocols
View specific protocols for Recombinant Mouse Fetuin A/AHSG Protein, CF (1563-PI):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Mouse Fetuin A/AHSG (rmFetuin A) (Catalog # 1563-PI)
- Recombinant Human Active Trypsin 3/PRSS3 (rhTrypsin 3) (Catalog # 3714-SE)
- Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhTrypsin 3 to 0.6 µg/mL in Assay Buffer.
- Prepare a curve of rmFetuin A (MW: 36651 Da) in Assay Buffer. Make the following serial dilutions: 6500, 1952, 651, 195.4, 65.1, 19.6, and 6.5 nM.
- Combine 20 µL of 0.6 µg/mL rhTrypsin 3, 100 µL of each dilution from the rmFetuin A curve, and 80 µL of Assay Buffer. Include a control (in duplicate) containing 180 µL of Assay Buffer and 20 µL of the diluted rhTrypsin 3.
- Incubate mixtures at room temperature for 30 minutes.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the incubated mixtures into a plate, and start the reaction by adding 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- Derive the 50% inhibition concentration (IC50) value for rmFetuin A by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
- The specific activity for rhTrypsin 3 at each point may be determined using the following formula (if needed):
|
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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
- rhTrypsin 3: 0.003 µg (1.17 nM)
- rmFetuin A curve: 1625, 488, 163, 48.9, 16.3, 4.9, and 1.625 nM
- Substrate: 10 µM
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