Recombinant Human Active Trypsin 3/PRSS3 Protein, CF

R&D Systems | Catalog # 3714-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Active Trypsin 3/PRSS3 Protein (3714-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

NP_002762

Structure / Form

Active form

Applications

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

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Trypsin 3/PRSS3 protein
Ile24-Ser247, with a C-terminal 10-His tag

Purity

>80%, 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

Ile24

Predicted Molecular Mass

26 kDa

SDS-PAGE

20-24 kDa, non-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 >4,000 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

3714-SE
Formulation Supplied as a 0.2 μm filtered solution in HCl, NaCl, CaCl2 and Brij-35.
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.

Background: Trypsin 3/PRSS3

Human Trypsin 3, encoded by the PRSS3 gene, is also known as mesotrypsin (1). Constituting less than 10% of the total trypsin content in normal pancreatic juice, it is one of the three trypsin isoforms produced by the pancreas (2). Compared to trypsin 1 and 2, one intriguing feature of Trypsin 3 is its resistance to polypeptide trypsin inhibitors, such as the Kunitz-type soybean trypsin inhibitor or the Kazal-type pancreatic secretory trypsin inhibitor. As revealed by the crystal structure, this resistance is likely due to the presence of an arginine residue in place of the highly conserved Gly198 (3). Trypsin 3 is synthesized in the pancreas and secreted into the duodenum lumen, where it is activated by enterokinase. One proposed physiological function of Trypsin 3 is degradation of trypsin inhibitors, which facilitates the digestion of those foods rich in these proteins (4).

References

  1. Nyaruhucha, 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_002762

Additional Trypsin 3/PRSS3 Products

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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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

For research use only

Related Research Areas

Serine Proteases and Regulators

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

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Protocols

View specific protocols for Recombinant Human Active Trypsin 3/PRSS3 Protein, CF (3714-SE):

Materials
  • Assay Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
  • Recombinant Human Active Trypsin 3/PRSS3 (rhTrypsin 3) (Catalog # 3714-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 rhTrypsin 3 to 0.06 µg/mL in Assay Buffer.
  2. Dilute Substrate to 80 µM in Assay Buffer.
  3. Load 50 µL of 0.06 µg/mL of rhTrypsin 3 into a plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 80 µM Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 405 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • rhTrypsin 3 (Active form): 0.003 µg
  • Substrate: 40 µM

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