Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Lys-ThioBenzyl ester (Z-Lys-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468. The specific activity is >75 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse Tryptase beta-1/MCP-7 protein
  Ile29-Phe273, with an C-terminal 10-His tag
• Accession #: Q02844
• N-terminal Sequence: Ile29
• Structure / Form: Mature form
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Tpsab1
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 28.8 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 36 kDa and 37 kDa, reducing conditions

Packaging, Storage & Formulations

• 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 opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl₂.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 50 mM MES, 1 M NaCl, pH 6.5
  • Recombinant Mouse Tryptase beta -1/MCPT-7 (rmTryptase beta -1) (Catalog # 1937-SE)
  • Substrate: Benzyloxcarbonyl Lys Thiobenzyl ester (Z-K-SBzl) (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
  • 96 well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmTryptase beta -1 to 10 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer containing 200 µM of DTNB.
  3. In a plate load 50 µL of 10 ng/µL rmTryptase beta -1, and start the reaction by adding 50 µL of the Substrate/DTNB mixture to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate Mixture.
  4. Read in kinetic mode for 15 minutes at an absorbance of 405 nm.
  5. Calculate specific activity using data from 1-15 minutes:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (OD/min) x well volume (L) x 10^12 pmol/mol) / (ext. coeff** (M^-1cm^-1) x path corr.*** (cm) x amount of enzyme (µg))

  
  *Adjusted for Substrate Blank
  **Using the extinction coefficient 13260 M^-1cm^-1
  ***Using the path correction 0.320 cm
  Note: the output of many spectrophotometers is in mOD Per Well:
  • rmTryptase beta -1: 0.5 µg
  • DTNB: 100 µM
  • Substrate: 100 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF

• Mast Cell Tryptase
• Mcp7
• MCP-7
• Mcpt7
• TPSB1
• TRYB1
• Tryptase beta1
• Tryptase beta-1

Background

Tryptase beta -1 is a serine protease with trypsin-like activity, which is sometimes also referred to as Mast Cell Protease 7 (1). It is stored in the secretory granules of mouse mast cells (2). It exhibits anticoagulant activity due to its ability to degrade fibrinogen in the presence of the diverse array of protease inhibitors in plasma (3). The deduced amino acid sequence for mouse Tryptase beta -1 consists of a signal peptide (residues 1-18), a pro region (residues 19-28) and the mature chain (residue 29‑273). The mature chain was expressed and purified.

1. McNeil, H.P. et al. (1992) Proc. Natl. Acad. Sci. USA 89:11174.
2. Matsumoto, R et al. (1995) J. Biol. Chem. 270:19524.
3. Huang, C. et al. (1997) J. Biol. Chem. 272:31885.

Publications for Tryptase beta-1/MCP-7/Mcpt7 (1937-SE)(4)

Publications using 1937-SE

• M Cecilia Berin Immunotherapy using algal-produced Ara h 1 core domain suppresses peanut allergy in mice Plant Biotechnol. J., 2016-01-23;14(7):1541-50. 2016-01-23 [PMID: 26801740] (ELISA (Standard), Mouse)
• Chen L, Schrementi M, Ranzer M, Wilgus T, DiPietro L Blockade of mast cell activation reduces cutaneous scar formation. PLoS ONE, 2014-01-22;9(1):e85226. 2014-01-22 [PMID: 24465509] (Bioassay, Mouse)
• de Souza DA, Toso VD, Campos MR, Lara VS, Oliver C, Jamur MC Expression of Mast Cell Proteases Correlates with Mast Cell Maturation and Angiogenesis during Tumor Progression. PLoS ONE, 2012-07-18;7(7):e40790. 2012-07-18 [PMID: 22815822] (Bioassay, Mouse)
• Bouchard J, Kim J, Beal D, Vaickus L, Craciun F, Remick D Acute oral ethanol exposure triggers asthma in cockroach allergen-sensitized mice. Am J Pathol, 2012-07-13;181(3):845-57. 2012-07-13 [PMID: 22796441] (ELISA (Standard))

Bioinformatics

• Gene Symbol: Tpsab1
• Uniprot:
  • Q02844()