Recombinant Mouse Matriptase/ST14 Catalytic Domain, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Mouse Matriptase/ST14 Catalytic Domain, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014). The specific activity is >4,000 pmol/min/µg, as measured under the described conditions.
Source
E. coli-derived mouse Matriptase/ST14 protein
Gly596-Val855, with an N-terminal Met and 6-His tag
The protein was auto-activated and further purified.
Accession #
N-terminal Sequence
Met & Val615
Protein/Peptide Type
Recombinant Enzymes
Gene
St14
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
3 kDa & 26 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
27 kDa (major) and minor auto-activation fragments, reducing conditions
Publications
Read Publications using
4735-SE in the following applications:

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 and Glycerol.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 50 mM NaCl, 0.01% (v/v) Tween® 20, pH 9.0
  • Recombinant Mouse Matriptase/ST14 Catalytic Domain (rmMatriptase) (Catalog # 4735-SE)
  • Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmMatriptase to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 50 µM in Assay Buffer.
  3. Load into a plate 50 µL of 0.2 ng/µL rmMatriptase, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 50 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
  5. 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 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A9891).

Per Well:
  • rmMatriptase: 0.010 µg
  • Substrate: 25 µM

Notes

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

Alternate Names for Recombinant Mouse Matriptase/ST14 Catalytic Domain, CF

  • EC 3.4.21
  • Epithin
  • HAI
  • Matriptase
  • Membrane-type serine protease 1
  • MTSP1
  • MT-SP1EC 3.4.21.109
  • Prostamin
  • PRSS14
  • Serine protease 14
  • Serine protease TADG-15
  • SNC19
  • SNC19MTSP1
  • ST14
  • suppression of tumorigenicity 14 (colon carcinoma)
  • suppression of tumorigenicity 14 (colon carcinoma, matriptase, epithin)
  • suppressor of tumorigenicity 14 protein
  • TADG15
  • TADG-15
  • TMPRSS14
  • tumor associated differentially expressed gene 15 protein
  • Tumor-associated differentially-expressed gene 15 protein

Background

Matriptase, a mouse type II membrane serine protease encoded by the ST14 (suppression of tumorigenicity 14) gene, is also known as epithin, and membrane-type serine protease 1/MT-SP1 (1-2). Its human ortholog MT-SP1/Matriptase (Catalog # 3946-SE), which shares 81% amino acid identity with epithin, has been thought to play an important role in tumor biology and is a potential target for anti-cancer therapy (3). Matriptase has a multidomain structure containing a putative N-terminal transmembrane region, two CUB domains, four LDLRA repeats, and a C-terminal serine protease domain (1). The protease domain of epithin starts with Val615. R&D Systems recombinant mouse Matriptase is an active protease and consists of the catalytic domain (Val615 to Val855) and a short peptide (Gly596 to Arg614).

  1. Cho, E. et al. (2001) J. Biol. Chem. 276:44581.
  2. List, K. et al. (2006) Mol. Med. 12:1.
  3. Uhland, K. (2006) Cell Mol. Life Sci. 63:2968.

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Publications for Matriptase/ST14 (4735-SE)(2)

We have publications tested in 2 confirmed species: Mouse, N/A.

We have publications tested in 2 applications: Enzyme Assay, Western Blot.


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Enzyme Assay
(1)
Western Blot
(1)
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(1)
N/A
(1)
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Bioinformatics

Gene Symbol St14
Uniprot