Tryptase gamma -1, also known as transmembrane tryptase, is a serine protease synthesized as a preproenzyme with a C-terminal transmembrane anchor (1, 2). Mast cells, mediators of inflammatory and allergic response, are found throughout the body concentrated near blood vessels in connective tissue and the mucous membranes of the respiratory and gastrointestinal tract (3). Upon activation, mast cells release granules that are enriched with neutral serine proteases including tryptases, chymase and cathepsin G (4). The recombinant mouse TPSG1 was expressed as a soluble protein terminated at residue 275, lacking the transmembrane domain, and corresponded to the proenzyme. The proenzyme can be cleaved by trypsin to form the active enzyme. The serine protease activity of trypsin-activated recombinant mouse TPSG1 can be inhibited by ecotin (Catalog # http://www.rndsystems.com/product_results.aspx?k=1328-PI">1328-PI). Typically, >95% protease activity is inhibited by ecotin at approximately a 10:1 molar ratio.
Recombinant Mouse Tryptase gamma-1/TPSG1 Protein, CF
R&D Systems | Catalog # 5517-SE
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Key Product Details
- R&D Systems CHO-derived Recombinant Mouse Tryptase gamma-1/TPSG1 Protein (5517-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
CHO
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived mouse Tryptase gamma-1/TPSG1 protein
His20-Ala275, with a C-terminal 10-His tag
Accession # Q9QUL7
His20-Ala275, with a C-terminal 10-His tag
Accession # Q9QUL7
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
His20
Predicted Molecular Mass
28 kDa
SDS-PAGE
37 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >100 pmol/min/μg, as measured under the described conditions.
The specific activity is >100 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
5517-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2. |
| 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.
|
Background: Tryptase gamma-1/TPSG1
References
- Wong, G.W. et al. (1999) J. Biol. Chem. 274:30784.
- Caughey, G.H. et al. (2000) J. Immunol. 164:6566.
- Harris, J.L. et al. (2001) J. Biol. Chem. 276:34941.
- Miller, H.R.P. and A.D. Pemberton (2002) Immunology 105:375.
Alternate Names
TPSG1, Tryptase gamma1
Entrez Gene IDs
25823 (Human)
Gene Symbol
TPSG1
UniProt
Additional Tryptase gamma-1/TPSG1 Products
Product Documents for Recombinant Mouse Tryptase gamma-1/TPSG1 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 Tryptase gamma-1/TPSG1 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Tryptase gamma-1/TPSG1 Protein, CF (5517-SE):
Materials
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Assay Buffer: 50 mM Tris, pH 8.5
- Recombinant Mouse Tryptase gamma ‑1/TPSG1 (rmTPSG1) (Catalog # 5517-SE)
- Trypsin (Sigma, Catalog # T-1426)
- Recombinant Human Serpin F2/ alpha 2‑Antiplasmin (rhSerpin F2) (Catalog # 1470-PI)
- Fluorogenic Peptide Substrate II: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(Dnp)-NH2 (Catalog # ES002), 2 mM in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmTPSG1 to 200 µg/mL in Activation Buffer.
- Dilute Trypsin to 0.164 µg/mL in Activation Buffer.
- Combine equal volumes of 200 µg/mL rmTPSG1 and 0.164 µg/mL Trypsin in a microfuge tube.
- Incubate at 37 °C for 30 min.
- Dilute rhSerpin F2 to 3.62 µg/mL in Assay Buffer.
- Add rhSerpin F2 to the reaction mixture making a ½ dilution for final concentrations of 1.81 µg/mL rhSerpin F2, 50 µg/mL rmTPSG1, and 0.041 µg/mL Trypsin.
- Incubate for 15 minutes at room temperature. Addition of rhSerpin F2 will stop the activity of Trypsin.
- Dilute rmTPSG1 to 2 ng/µL in Assay Buffer.
- Dilute fluorogenic peptide Substrate to 20 µM in Assay Buffer.
- Load into plate 50 µL of the diluted samples and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
- 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).
Per Well:
- rmTPSG1: 0.100 µg
- Substrate: 10 µM
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