Tryptases are trypsin-like serine proteases, with beta tryptases as the main isoenzymes expressed in mast cells (1). They are stored in secretory granules of mast cells, where they form active tetramers with heparin proteoglycan. Because of the unique arrangement of the active sites in the tetramer, which are facing a narrow central pore, beta tryptases are resistant to macromolecule protease inhibitors (2). When mast cells are activated, beta tryptases are released along with other proteins in secretory granules, participating in provoking inflammatory conditions (3). beta tryptases have been implicated as mediators in the pathogenesis of asthma and other allergic disorders. According to the protein sequence, this product was previously labeled as human Tryptase beta-2 protein upon release. This product has since been renamed as Recombinant Human Tryptase alpha/beta-1 to reflect current sequence alignment per sequence revisions and classification adjustments that have occurred in the NCBI databases.
Recombinant Human Tryptase alpha/beta-1 Protein, CF
R&D Systems | Catalog # 3796-SE
Key Product Details
- R&D Systems NS0-derived Recombinant Human Tryptase alpha/beta-1 Protein (3796-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
Accession Number
Structure / Form
Applications
Product Specifications
Source
Met1-Pro275, with a C-terminal 10-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >650 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3796-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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.
|
Background: Tryptase alpha/beta-1
References
- Caughey, G. H. 2004, in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. pp. 1535.
- Sommerhoff, C.P. et al. (1999) Proc. Natl. Acad. Sci. USA. 96:10984.
- Hallgren, J. and G. Pejler (2006) FEBS J. 273:1871.
Alternate Names
Gene Symbol
UniProt
Additional Tryptase alpha/beta-1 Products
Product Documents for Recombinant Human Tryptase alpha/beta-1 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 Human Tryptase alpha/beta-1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Tryptase alpha/beta-1 Protein, CF
Powered by Bioz
Customer Reviews for Recombinant Human Tryptase alpha/beta-1 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Tryptase alpha/beta-1 Protein, CF and earn rewards!
Have you used Recombinant Human Tryptase alpha/beta-1 Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human Tryptase alpha/beta-1 Protein, CF (3796-SE):
- Maturation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Heparin Incubation Buffer (HIB): 100 µg/mL Heparin (Sigma, Catalog # H3393), 50 mM MES, pH 5.5
- Assay Buffer: 50 mM Tris, pH 8.5
- Recombinant Human Tryptase alpha/beta-1 (rhTPSAB1) (Catalog # 3796-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
- 1, 10 Phenanthroline (Sigma, Catalog # 320056)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhTPSAB1 to 100 µg/mL in Maturation Buffer containing Thermolysin at a final concentration of 0.1 µg/mL.
- Incubate at room temperature for 15 minutes.
- Stop Thermolysin activity by adding an equal volume of 10 mM 1, 10 Phenanthroline prepared in HIB.
- Dilute the matured rhTPSAB1 to 10 µg/mL in HIB.
- Incubate for 2 hours at room temperature.
- Dilute 10 µg/mL rhTPSAB1 to 2.5 µg/mL in HIB.
- Dilute 2.5 µg/mL rhTPSAB1 to 0.50 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of 0.50 µg/mL rhTPSAB1 into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), 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:
- rhTPSAB1: 0.025 µg
- Substrate: 10 µM