Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF

R&D Systems | Catalog # 1937-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Tryptase beta-1/Mcpt7 Protein (1937-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

Q02844

Structure / Form

Mature form

Applications

Enzyme Activity
View all Tryptase beta-1/MCP-7 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Tryptase beta-1/MCP-7 protein
Ile29-Phe273, with an C-terminal 10-His tag

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

Ile29

Predicted Molecular Mass

28.8 kDa

SDS-PAGE

36 kDa and 37 kDa, reducing conditions

Activity

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.

Formulation, Preparation, and Storage

1937-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Tryptase beta-1/MCP-7

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.

References

  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.

Long Name

Mast Cell Protease 7

Alternate Names

Mast Cell Tryptase, MCP-7, Mcp7, Mcpt7, TPSB1, TRYB1, Tryptase beta1

Entrez Gene IDs

100503895 (Mouse)

Gene Symbol

TPSAB1

UniProt

Q02844

Additional Tryptase beta-1/MCP-7 Products

Product Documents for Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF

For research use only

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

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Protocols

View specific protocols for Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF (1937-SE):

Materials
  • 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 1012 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

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