Recombinant Human Tryptase epsilon/BSSP-4 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

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 #
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

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 (MP Biochemicals, Catalog # SB001)
  • 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 1012 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.

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Bioinformatics

Gene Symbol PRSS22
Uniprot