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Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, CF

Catalog #: 1937-SE
4 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
1937-SE-020
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 Mouse Tryptase beta-1/Mcpt7 Protein, CF Summary

Product Specifications

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.
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.
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
Analysis
Ile29
Structure / Form
Mature form
Predicted Molecular Mass
28.8 kDa
SDS-PAGE
36 kDa and 37 kDa, reducing conditions

Product Datasheets

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1937-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.

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.

Assay Procedure

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
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: 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
Entrez Gene IDs
100503895 (Mouse)
Alternate Names
Mast Cell Tryptase; Mcp7; MCP-7; Mcpt7; TPSB1; TRYB1; Tryptase beta1; Tryptase beta-1

Citations for Recombinant Mouse Tryptase beta-1/Mcpt7 Protein, 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. Immunotherapy using algal-produced Ara h 1 core domain suppresses peanut allergy in mice
    Authors: M Cecilia Berin
    Plant Biotechnol. J., 2016-01-23;14(7):1541-50.
    Species: Mouse
    Sample Types: Protein
    Applications: ELISA (Standard)
  2. Blockade of mast cell activation reduces cutaneous scar formation.
    Authors: Chen L, Schrementi M, Ranzer M, Wilgus T, DiPietro L
    PLoS ONE, 2014-01-22;9(1):e85226.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Expression of Mast Cell Proteases Correlates with Mast Cell Maturation and Angiogenesis during Tumor Progression.
    Authors: de Souza DA, Toso VD, Campos MR, Lara VS, Oliver C, Jamur MC
    PLoS ONE, 2012-07-18;7(7):e40790.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Acute oral ethanol exposure triggers asthma in cockroach allergen-sensitized mice.
    Authors: Bouchard J, Kim J, Beal D, Vaickus L, Craciun F, Remick D
    Am J Pathol, 2012-07-13;181(3):845-57.
    Applications: ELISA (Standard)

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