Mast Cell Protease-11 (MCP-11) is encoded by Prss34, one of 13 genes on mouse chromosome 17A3.3 that correspond to functional trypsin-like serine proteases (1). The deduced amino acid sequence of mouse MCP-11 consists of 318 residues with a signal peptide (residues 1 to 19), a pro region (residue 20 to 34), and a catalytic domain (35 to 318). The mRNA is preferentially expressed in spleen and bone marrow. The mouse MCP-11 (residues 20 to 318) was expressed in the NS0 cells with a foreign signal peptide. After being treated with thermolysin, the purified enzyme is active against a peptide substrate described in the Activity Assay Protocol. Apparently, the human gene corresponding to Prss34 encodes a protein that is not enzymatically active due to a mutation that leads to a premature translation termination codon.
Recombinant Mouse Mast Cell Protease-11/Prss34 Protein, CF
R&D Systems | Catalog # 2857-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Mast Cell Protease-11/Prss34 Protein (2857-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
Structure / Form
Pro form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Mast Cell Protease-11/Prss34 protein
Met20-Ser318, with a C-terminal 10-His tag
Met20-Ser318, with a 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
Met20
Predicted Molecular Mass
34 kDa
SDS-PAGE
48 kDa doublet, reducing conditions
Activity
Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Arg-ThioBenzyl ester (Z-R-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 >20,000 pmol/min/µg, as measured under the described conditions.
The specific activity is >20,000 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2857-SE
| Formulation | Supplied as a 0.2 μm filtered solution in MES 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: Mast Cell Protease-11/Prss34
References
- Wong, G.W. et al. (2004) J. Biol. Chem. 279:2438.
Alternate Names
Mast Cell Protease11, MCP-11, MCPT11, Prss34
Entrez Gene IDs
328780 (Mouse)
Gene Symbol
PRSS34
UniProt
Additional Mast Cell Protease-11/Prss34 Products
Product Documents for Recombinant Mouse Mast Cell Protease-11/Prss34 Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Mast Cell Protease-11/Prss34 Protein, CF
For research use only
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Protocols
View specific protocols for Recombinant Mouse Mast Cell Protease-11/Prss34 Protein, CF (2857-SE):
Materials
- Activation Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, pH 7.5 (TCN)
- Assay Buffer: 50 mM Tris, pH 8.0
- Recombinant Mouse Mast Cell Protease‑11/Prss34 (rmMCP-11) (Catalog # 2857-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
- Substrate: Z-Arg-SBzl (SM Biochemicals, Catalog # SMSB01), 10 mM in DMSO
- 5,5'Dithio-bis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130)
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rmMCP-11 to 200 µg/mL with Activation Buffer.
- Dilute Thermolysin to 0.4 µg/mL with Activation Buffer.
- Mix equal volumes of 200 µg/mL rmMCP-11 and 0.4 µg/mL Thermolysin for final concentrations of 50 µg/mL and 0.2 µg/mL, respectively.
- Incubate at 37 °C for 30 minutes.
- Stop the reaction with 10 mM 1,10 Phenanthroline.
- Dilute activated rmMCP-11 to 0.1 ng/µL in Assay Buffer.
- Dilute substrate to 200 µM in Assay Buffer with 200 µM of DTNB.
- Load 50 µL of the 0.1 ng/µL rmMCP-11 into plate, 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 mix without any rmMCP-11.
- Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
- Calculate specific activity:
|
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 0.32 cm
Note: the output of many spectrophotometers is in mOD Per Well:
- rmMCP-11: 0.005 µg
- DTNB: 100 µM
- Substrate: 100 µM
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