Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF

R&D Systems | Catalog # 7235-SE

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Mouse Active Trypsin 3/PRSS3 Protein (7235-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

E. coli

Accession Number

NP_035775

Applications

Enzyme Activity
View all Trypsin 3/PRSS3 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived mouse Trypsin 3/PRSS3 protein
Ile24-Asn246, with an N-terminal Met, 6-His tag and GGSGSGGGSSR linker

Purity

>85%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

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

N-terminal Sequence Analysis

Ile24

Predicted Molecular Mass

24 kDa

SDS-PAGE

24-27 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >30,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7235-SE
Formulation Supplied as a 0.2 μm filtered solution in Citrate, NaCl, CaCl2, Benz, DTT, and Glycerol.
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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Trypsin 3/PRSS3

Mouse Trypsin 3, encoded by the Prss3 gene, is also known as mesotrypsin and brain trypsin (1). It is one of three trypsin isoforms produced by the pancreas (2). It is secreted into the duodenum lumen, where it is activated by enterokinase. Trypsin 3 differs from Trypsin 1 and 2 because it is resistant to polypeptide trypsin inhibitors such as the Kunitz-type soybean trypsin inhibitor and the Kazal‑type pancreatic secretory trypsin inhibitor (3). It has been proposed that a function of Trypsin 3 is the degradation of trypsin inhibitors to facilitate the digestion of foods rich in these proteins (4). Recombinant mouse Trypsin 3 is expressed and processed from E.coli, which yields the purified mature chain of mouse Trypsin 3.

References

  1. Nyarahucha, C.N.M. et al. (1997) J. Biol. Chem. 272:10573.
  2. Rinderknecht, H. et al. (1984) Gastroenterology 86:681.
  3. Katona, G. et al. (2002) J. Mol. Biol. 315:1209.
  4. Szmola, R. et al. (2003) J. Biol. Chem. 278:48580.

Alternate Names

Mesotrypsin, MTG, PRSS3, PRSS4, Trypsin 4, Trypsinogen 15

Entrez Gene IDs

5646 (Human); 22073 (Mouse); 362347 (Rat)

Gene Symbol

PRSS3

UniProt

NP_035775

Additional Trypsin 3/PRSS3 Products

Product Documents for Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF

For research use only

Related Research Areas

Serine Proteases and Regulators

Citations for Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF (7235-SE):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij35, pH 8.0
  • Recombinant Mouse Active Trypsin 3/PRSS3 (rmTrypsin 3) (Catalog # 7235-SE)
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys-(DNP)-NH2(Catalog # ES002), 2 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 rmTrypsin 3 to 0.007 ng/μL in Assay Buffer.
  2. Dilute Substrate to 20 μM in Assay Buffer.
  3. Load into a plate 50 μL of 0.007 ng/μL rmTrypsin 3, and start the reaction by adding 50 μL of 20 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 20 μM Substrate.
  4. Read plate at excitation and emission wavelengths of 320 nm and 405 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • rmTrypsin 3: 0.35 ng (0.00035 μg)
  • Substrate: 10 μM

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