Recombinant Human Tryptase epsilon/BSSP-4 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human Tryptase epsilon/BSSP-4 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave benzyloxycarbonyl-Arg-thiobenzyl ester (Z-R-SBzl). The specific activity is >5,000 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Tryptase epsilon/BSSP-4 protein
  Ala33-Ser317 & Val50-Ser317, both with a C-terminal 10-His tag
• Accession #: Q9GZN4
• N-terminal Sequence: Ala33 & Val50
• Structure / Form: Pro and mature forms
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PRSS22
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 32 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: Multiple bands between 33-38 kDa, reducing conditions

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 Sodium Acetate, NaCl and Glycerol.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 0.05% (w/v) Brij-35, pH 8.0
  • Recombinant Human Tryptase epsilon /BSSP‑4 (rhBSSP-4) (Catalog # 5047-SE)
  • Substrate: Z-Arg-SBzl (SM Biochemicals, Catalog # SMSB01)
  • DTNB (5,5’-dithibis(2-nitrobenzoic acid)) (Sigma, Catalog # D-8130)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhBSSP-4 to 0.5 ng/µL in Aassay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer containing 200 µM DTNB.
  3. Load 50 µL of 0.5 ng/µL rhBSSP-4 into a plate, and start the reaction by adding 50 µL of 200 µM Substrate to wells. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  4. Read at an absorbance of 405 nm for 5 minutes in kinetic mode.
  5. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (OD/min) x well volume (L) x 10^12 pmol/M) / (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:
  • rhBSSP-4: 0.025 µg
  • Substrate: 100 µM

Notes

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

Alternate Names for Recombinant Human Tryptase epsilon/BSSP-4 Protein, CF

• BSP-2
• BSSP4
• BSSP-4
• hBSSP-4
• MGC9599
• prosemin
• protease, serine S1 family member 22
• protease, serine, 22
• PRSS22
• serine protease 22
• serine protease 26
• SP001LA
• Tryptase epsilon
• UNQ302/PRO343

Background

Tryptase epsilon , also known as brain-specific protease-4 (BSSP-4) and prosemin, is expressed in the brain and epithelium-rich tissues such as the lung and eye (1, 2). BSSP-4 is a serine protease related to trypsin, preferentially hydrolyzing substrates after arginine and lysine residues. However, Tryptase epsilon is less susceptible to inhibition by common trypsin inhibitors such as aprotinin, alpha 1-antitrypsin and secretory leukocyte protease inhibitor (3). Tryptase  epsilon efficiently converts pro‑urokinase‑type plasminogen activator to its mature, active form (3).

1. Wong, G.W. et al. (2004) J. Biol. Chem. 279:2438.
2. Wong, G.W. et al. (2001) J. Biol. Chem. 276:49169.
3. Yasuda, S. et al. (2005) Blood 105:3893.

Bioinformatics

• Gene Symbol: PRSS22
• Uniprot:
  • Q9GZN4()