Home / Airway Trypsin-like Protease/HAT / Recombinant Human Airway Trypsin-like Protease/HAT, CF

Recombinant Human Airway Trypsin-like Protease/HAT, CF

Catalog #: 2695-SE
4 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
2695-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (4)
FAQs
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Airway Trypsin-like Protease/HAT, CF Summary

Product Specifications

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.
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.
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.
Accession #
O60235
N-terminal Sequence
Analysis
Ile187 & His
Structure / Form
Active
Predicted Molecular Mass
25 kDa & 17 kDa  
SDS-PAGE
27 kDa and 18 kDa, reducing conditions

Product Datasheets

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2695-SE

Product Datasheet
COA
SDS

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.

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Assay Procedure

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
  1. Dilute rhHAT to 0.02 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. 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.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively in kinetic mode for 5 minutes.
  5. 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).

Per Well:
  • rhHAT: 0.001 µg
  • Substrate: 100 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Airway Trypsin-like Protease/HAT

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.

References
  1. Yasuoka, S. et al. (1997) Am. J. Respir. Cell Mol. Biol. 16:300.
  2. Yamaoka, K. et al. (1998) J. Biol. Chem. 273:11895.
  3. Hooper, J.D. et al. (2001) J. Biol. Chem. 276:857.
  4. Iwakiri, K. et al. (2004) J. Invest. Dermatol. 122:937.
  5. Chokki, M. et al. (2004) Am. J. Respir. Cell Mol. Biol. 30:470.
Long Name
Human Airway Trypsin-like Protease
Entrez Gene IDs
9407 (Human); 231382 (Mouse)
Alternate Names
airway trypsin like protease; Airway trypsin-like protease; EC 3.4.21; EC 3.4.21.-; HAT; MGC150587; MGC150588; TMPRSS11D; transmembrane protease serine 11D; transmembrane protease, serine 11D

Citations for Recombinant Human Airway Trypsin-like Protease/HAT, 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.

4 Citations: Showing 1 - 4
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  1. Airway proteolytic control of pneumococcal competence
    Authors: Echlin, H;Iverson, A;Sardo, U;Rosch, JW;
    PLoS pathogens
    Species: Human
    Sample Types: Peptide
    Applications: Bioassay
  2. A peptide-based approach to evaluate the adaptability of influenza A virus to humans based on its hemagglutinin proteolytic cleavage site
    Authors: MR Straus, GR Whittaker
    PLoS ONE, 2017-03-30;12(3):e0174827.
    Species: Virus - Influenza A
    Sample Types: Peptide
    Applications: Enzyme Assay
  3. TMPRSS2 and ADAM17 cleave ACE2 differentially and only proteolysis by TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein.
    Authors: Heurich A, Hofmann-Winkler H, Gierer S, Liepold T, Jahn O, Pohlmann S
    J Virol, 2013-11-13;88(2):1293-307.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  4. Impact of host proteases on reovirus infection in the respiratory tract.
    Authors: Nygaard RM, Golden JW, Schiff LA
    J. Virol., 2011-11-09;86(2):1238-43.
    Species: Virus
    Sample Types: Virus
    Applications: Enzyme Assay

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