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Recombinant Human Matriptase/ST14 Catalytic Domain, CF

R&D Systems | Catalog # 3946-SEB

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Matriptase/ST14 Catalytic Domain (3946-SEB)
  • 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

E. coli

Accession Number

NP_068813

Applications

Enzyme Activity
View all Matriptase/ST14 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Matriptase/ST14 protein
Gly596-Val855, with an N-terminal Met and 6-His tag
The protein was purified, auto-activated and further purified.

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Val615

Predicted Molecular Mass

26 kDa

SDS-PAGE

27 kDa (major) and minor auto-activation fragments, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014).
The specific activity is >10,000 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 4 reviews rated 5 using 3946-SEB in the following applications:

Scientific Data Images for Recombinant Human Matriptase/ST14 Catalytic Domain, CF

Recombinant Human Matriptase/ST14 Catalytic Domain Enzyme Activity

Recombinant Human Matriptase/ST14 Catalytic Domain Enzyme Activity

Recombinant Human Matriptase is measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014).

Formulation, Preparation, and Storage

3946-SEB
Formulation Supplied as a 0.2 μm filtered solution in Tris-HCl and Glycerol.
Shipping The product is shipped with polar packs. 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: Matriptase/ST14

Human matriptase, encoded by the ST14 (suppression of tumorogenicity 14) gene, is also known as tumor associated differentially expressed gene 15 protein/TADG‑15), epithin, and membrane‑type serine protease 1/MT‑SP1 (1). Predicted to have a significant role in tumor biology, matriptase may be a novel target for anti‑cancer therapy (2). However, expressed in most human epithelia, matriptase is also important in several physiological processes (1). For example, it activates prostasin to initiate a protease cascade that is essential for epidermal differentiation (3), and it converts a single‑chain IGFBP-rp1 into the two‑chain form (4). Matriptase is a type II transmembrane serine protease with a complex modular structure (1). The 855 amino acid (aa) sequence of human matriptase consists of a cytoplasmic tail (aa 1‑55), a transmembrane domain (aa 56‑76), and an extracellular portion (aa 77‑855). The latter contains the following domains: SEA (aa 86‑201), two CUBs (aa 214‑334 and 340‑447), four LDLRAs (aa 452‑486, 487‑523, 524‑560, and 566‑603), and a serine protease (aa 615‑855). The physiological activation of the single‑chain zymogen requires the cleavage at the SEA domain within the ER or Golgi, association with HAI-1, which facilitates the transport of the protease to the cell surface, and auto‑cleavage at QAR-V(615)VGG (1). The activated matriptase is inhibited by HAI-1, and the resulting HAI-1 complex can be shed from the cell surface (1). R&D Systems recombinant human (rh) ST14 corresponds to the catalytic domain, and is inhibited effectively by rhHAI-1 and rhHAI-2A (Catalog # http://www.rndsystems.com/product_results.aspx?k=1048-PI">1048‑PI and http://www.rndsystems.com/product_results.aspx?k=1106-PI">1106‑PI).

References

  1. List, K. et al. (2006) Mol. Med. 12:1.
  2. Uhland, K. (2006) Cell. Mol. Life Sci. 63:2968.
  3. Netzel-Arnett, S. et al. (2006) J. Biol. Chem. 281:32941.
  4. Ahmed, S. et al. (2006) FEBS J. 273:615.

Alternate Names

Epithin, MTSP1, PRSS14, SNC19, ST14, TADG-15, TMPRSS14

Entrez Gene IDs

6768 (Human); 19143 (Mouse); 114093 (Rat); 102117178 (Cynomolgus Monkey)

Gene Symbol

ST14

UniProt

NP_068813

Additional Matriptase/ST14 Products

Product Documents for Recombinant Human Matriptase/ST14 Catalytic Domain, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Matriptase/ST14 Catalytic Domain, CF

For research use only

Citations for Recombinant Human Matriptase/ST14 Catalytic Domain, CF

Customer Reviews for Recombinant Human Matriptase/ST14 Catalytic Domain, CF (4)

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  • Recombinant Human Matriptase/ST14 Catalytic Domain, CF
    Name: Anonymous
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 12/11/2019
    Substrate fusion proteins were purified from Drosophila S2R+ cells using anti-RFP magnetic agarose Substrates were incubated with 0.1 ug matriptase at 37°C for one hour. Reaction was stopped by reducing/denaturing conditions. Samples were examined by SDS-PAGE/western blots and antibody stainings using respective antibodys.
    Recombinant Human Matriptase/ST14 Catalytic Domain, CF 3946-SEB
  • Recombinant Human Matriptase/ST14 Catalytic Domain, CF
    Name: Anonymous
    Application: SDS-PAGE Control
    Verified Customer | Posted 04/07/2019
    Recombinant Human Matriptase/ST14 Catalytic Domain, CF 3946-SEB
  • Recombinant Human Matriptase/ST14 Catalytic Domain, CF
    Name: Anonymous
    Application: SDS-PAGE Control
    Verified Customer | Posted 03/07/2019
    Protein sample treated with Matriptase enzyme was cleaved after 30min incubation compared with control (untreated). Protein ladder ran to compare sizes
    Recombinant Human Matriptase/ST14 Catalytic Domain, CF 3946-SEB
  • Name: Anonymous
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 07/31/2018
    5nM of activated ST14 used
    Recombinant Human Matriptase/ST14 Catalytic Domain, CF 3946-SEB

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Protocols

View specific protocols for Recombinant Human Matriptase/ST14 Catalytic Domain, CF (3946-SEB):

Materials
  • Assay Buffer: 50 mM Tris, 50 mM NaCl, 0.01% (v/v) Tween® 20, pH 9.0
  • Recombinant Human Matriptase/ST14 Catalytic Domain (rhMatriptase) (Catalog # 3946-SEB)
  • Substrate:  BOC-Gln-Ala-Arg-AMC (Catalog # ES014), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMatriptase to 0.1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 50 µM in Assay Buffer.
  3. Load 50 µL of 0.1 µg/mL rhMatriptase into a plate, 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 Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), 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 (AMC) (Sigma, Catalog # A9891).

Per Well:

  • rhMatriptase: 0.005 µg
  • Substrate: 25 µM

FAQs for Recombinant Human Matriptase/ST14 Catalytic Domain, CF

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  • Q: Catalog # 3946-SEB is formulated with 10% glycerol. Will the glycerol prevent it from freezing?

    A: No, this protein in 10% glycerol will freeze at -20 C and -80 C. 

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