Recombinant Human Tryptase beta-2 Protein, CF

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Recombinant Human Tryptase beta-2 Protein, CF Summary

Product Specifications

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002). The specific activity is >650 pmol/min/µg, as measured under the described conditions. 
Mouse myeloma cell line, NS0-derived human Tryptase beta-2/TPSB2 protein
Met1-Pro275, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Structure / Form
Pro form
Predicted Molecular Mass
29 kDa
38 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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.

Assay Procedure

  • Maturation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Heparin Incubation Buffer (HIB): 100 µg/mL Heparin (Sigma, Catalog # H3393), 50 mM MES, pH 5.5
  • Assay Buffer: 50 mM Tris, pH 8.5
  • Recombinant Human Tryptase  beta ‑2/TPSB2 (rhTPSB2) (Catalog # 3796-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
  • 1, 10 Phenanthroline (Sigma, Catalog # 320056)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhTPSB2 to 100 µg/mL in Maturation Buffer containing Thermolysin at a final concentration of 0.1 µg/mL.
  2. Incubate at room temperature for 15 minutes.
  3. Stop Thermolysin activity by adding an equal volume of 10 mM 1, 10 Phenanthroline prepared in HIB.
  4. Dilute the matured rhTPSB2 to 10 µg/mL in HIB.
  5. Incubate for 2 hours at room temperature.
  6. Dilute 10 µg/mL rhTPSB2 to 2.5 µg/mL in HIB.
  7. Dilute 2.5 µg/mL rhTPSB2 to 0.50 µg/mL in Assay Buffer.
  8. Dilute Substrate to 20 µM in Assay Buffer.
  9. Load 50 µL of 0.50 µg/mL rhTPSB2 into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  10. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  11. 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:
  • rhTPSB2: 0.025 µg
  • Substrate: 10 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.


Background: Tryptase beta-2/TPSB2

Tryptases are trypsin-like serine proteases, and beta  tryptases appear to be the main isoenzymes expressed in mast cells (1). They are stored in secretory granules of mast cells, where they form active tetramers with heparin proteoglycan. Because of the unique arrangement of the active sites in the tetramer, which are facing a narrow central pore, beta  tryptases are resistant to macromolecule protease inhibitors (2). When mast cells are activated, beta  tryptases are released along with other proteins in secretory granules, participating in provoking inflammatory conditions (3). beta  tryptases have been implicated as mediators in the pathogenesis of asthma and other allergic disorders.

  1. Caughey, G. H. 2004, in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. pp. 1535.
  2. Sommerhoff, C.P. et al. (1999) Proc. Natl. Acad. Sci. USA. 96:10984.
  3. Hallgren, J. and G. Pejler (2006) FEBS J. 273:1871.  
Entrez Gene IDs
64499 (Human); 17229 (Mouse); 29268 (Rat)
Alternate Names
EC 3.4.21; EC; mast cell tryptase beta II; mast cell tryptase beta III; TPS2; TPS2tryptase beta-2; TPSB2; tryptase beta 2 (gene/pseudogene); tryptase beta 2; tryptase beta II; tryptase beta III; Tryptase beta2; Tryptase beta-2; Tryptase II; tryptase III; tryptase-2; tryptaseB; tryptaseC; Tryptase-C


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