HAT was initially purified from the sputum of patients with chronic airway diseases (1). Subsequent cloning revealed it as a member type II transmembrane serine proteases (2, 3). HAT has been shown to induce PAR-2 mediated IL-8 release in psoriasis vulgaris and increase mucin expression in airway epithelial cells (4, 5). Located in the cells of the submucosal serous glands of the bronchi and trachea, the isolated enzyme had the N-terminal sequence of ILGGTEAEEG, which corresponded to the start of the C-terminal catalytic domain (residues 187 to 418) of the deduced sequence (1, 2). The N-terminal region consisted of a short cytoplasmic tail (residues 1 to 20), a transmembrane domain (residues 21 to 41), and a SEA domain (residues 44 to 164). The ectodomain of HAT is expressed and the active enzyme is purified.
Recombinant Human Airway Trypsin-like Protease/HAT, CF
R&D Systems | Catalog # 2695-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Airway Trypsin-like Protease/HAT (2695-SE)
- 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
Structure / Form
Active
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Airway Trypsin-like Protease/HAT protein
Ala42-Ile418 with an N-terminal 6-His tag
The protein was purified, activated and further purified.
Ala42-Ile418 with an N-terminal 6-His tag
The protein was purified, activated and further purified.
Purity
>90%, 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
Ile187 & His
Predicted Molecular Mass
25 kDa & 17 kDa
SDS-PAGE
27 kDa and 18 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, t-butoxycarbonyl-Phe-Ser-Arg-7-amino-4-methyl coumarin (Boc-FSR-AMC).
The specific activity is >50,000 pmol/min/µg, as measured under the described conditions.
The specific activity is >50,000 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2695-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 25 mM Tris, 150 mM NaCl and 0.05% Brij-35, 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: Airway Trypsin-like Protease/HAT
References
- Yasuoka, S. et al. (1997) Am. J. Respir. Cell Mol. Biol. 16:300.
- Yamaoka, K. et al. (1998) J. Biol. Chem. 273:11895.
- Hooper, J.D. et al. (2001) J. Biol. Chem. 276:857.
- Iwakiri, K. et al. (2004) J. Invest. Dermatol. 122:937.
- Chokki, M. et al. (2004) Am. J. Respir. Cell Mol. Biol. 30:470.
Long Name
Human Airway Trypsin-like Protease
Alternate Names
TMPRSS11D
Gene Symbol
TMPRSS11D
UniProt
Additional Airway Trypsin-like Protease/HAT Products
Product Documents for Recombinant Human Airway Trypsin-like Protease/HAT, 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 Human Airway Trypsin-like Protease/HAT, CF
For research use only
Related Research Areas
Citations for Recombinant Human Airway Trypsin-like Protease/HAT, CF
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Protocols
View specific protocols for Recombinant Human Airway Trypsin-like Protease/HAT, CF (2695-SE):
Materials
- Assay Buffer: 50 mM Tris, 0.05% (w/v) Brij-35, pH 9.5
- Recombinant Human Airway Trypsin‑like Protease/HAT (rhHAT) (Catalog # 2695-SE)
- Substrate: t-Butyloxycarbonyl Phe-Ser Arg 7-amino-4-methyl coumarin (Bachem, Catalog # I-1400), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhHAT to 0.02 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- In a plate load 50 µL of 0.02 ng/µL rhHAT, and start the reaction by adding 50 µL of 200 µM Substrate to wells. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 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 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).
- rhHAT: 0.001 µg
- Substrate: 100 µM