Recombinant Mouse HGF Activator Protein, CF Summary
Product Specifications
Met1-Ser653 with a C-terminal 10-His tag
The pro form was purified, activated and further purified.
Analysis
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.
1200-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2. |
| Shipping | The product is shipped with dry ice or equivalent. 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, 0.01% CHAPS, pH 9.0
- Recombinant Mouse HGFA (rmHGFA) (Catalog # 1200-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 rmHGFA to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of 2 ng/µL rmHGFA in a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank by combining 50 µL of 20 µM Substrate with 50 µL of Assay Buffer.
- Read at excitation and emission wavelengths of 320 nm and 405 nm, 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
- rmHGFA: 0.100 µg]
- Substrate: 10 µM
Reconstitution Calculator
Background: HGF Activator
Hepatocyte Growth Factor Activator (HGFA) is a serine endopeptidase that cleaves at the peptide bond between Arg494 and Val495 of single-chain human HGF precursor, generating the active heterodimer (1). HGFA is produced and secreted by the liver and normally circulates in the blood as an inactive zymogen (2, 3). The zymogen has a weak affinity for heparin but acquires a strong affinity for heparin upon activation that is linked to blood coagulation. This property may ensure the local action of this enzyme at the site of tissue injury (3). Mouse HGFA precursor (653 amino acid residues) contains several predicted domains including a signal peptide (residues 1‑29), a propeptide (residues 30‑369), and a mature and active form (residues 370 to 653) that is further processed into a short chain (residues 370‑405) and a long chain (residues 406‑653). The short chain and the long chain (catalytic domain) may form a disulfide bond linked dimer. HGFA can be activated by autocatalysis or by thrombin (4). The active protease can be inhibited by HGFA inhibitors (HAIs). Two HAIs, HAI-1 and HAI-2, are known in mouse and human. HAI-1 is not only an inhibitor, but also a specific acceptor of active HGFA, acting as a reservoir of this enzyme on the cell surface (5).
- Kitamura, N. (2004) in Handbook of Proteolytic Enzymes (Barrett, A.J. et al. Eds.), p. 1712, Academic Press, San Diego.
- Miyazawa, K. et al. (1993) J. Biol. Chem. 268:10024.
- Miyazawa, K. et al. (1996) J. Biol. Chem. 271:3615.
- Shimomura, T. et al. (1993) J. Biol. Chem. 268:22927.
- Kataoka, H. et al. (2000) J. Biol. Chem. 275:40453.
Citations for Recombinant Mouse HGF Activator 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.
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The mitochondrial protein OPA1 regulates the quiescent state of adult muscle stem cells
Authors: N Baker, S Wade, M Triolo, J Girgis, D Chwastek, S Larrigan, P Feige, R Fujita, C Crist, MA Rudnicki, Y Burelle, M Khacho
Cell Stem Cell, 2022-08-22;29(9):1315-1332.e9.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo -
Circulating healing (CH) cells expressing BST2 are functionally activated by the injury-regulated systemic factor HGFA
Authors: C Lo Sicco, D Reverberi, F Villa, U Pfeffer, R Quarto, R Cancedda, R Tasso
Stem Cell Res Ther, 2018-11-08;9(1):300.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo
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