Recombinant Human Kallikrein 2 Protein, CF

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

Recombinant Human Kallikrein 2 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a flourogenic peptide substrate Pro-Phe-Arg-7-amido-4-methylcoumarin (PFR-AMC). The specific activity is >125 pmol/min/μg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Kallikrein 2 protein
  Met1-Pro261, with a C-terminal 10-His tag
• Accession #: P20151
• N-terminal Sequence: Ile25 & Leu124
• Protein/Peptide Type: Recombinant Enzymes
• Gene: KLK2
• Purity: >85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 28 kDa & 16 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: 34 kDa & 15-18 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Sodium Citrate and NaCl.
• Purity: >85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl₂, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human Kallikrein 2 (rhKLK2) (Catalog # 4104-SE)
  • Substrate: Pro-Phe-Arg-AMC (Bachem, Catalog # I-1295), 10 mM stock in DMSO
  • 96-well black plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhKLK2 to 4 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. In a plate, load 50 µL of 4 µg/mL rhKLK2, and start the reaction by adding 50 µL of the 200 µM Substrate. As a Substrate Blank combine 50 µL of Assay Buffer with 50 µL of the 200 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 0 to 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 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

  Per Well:
  • rhKLK2: 0.2 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 Kallikrein 2 Protein, CF

• EC 3.4.21
• EC 3.4.21.35
• glandular kallikrein 2
• Glandular kallikrein-1
• hGK-1
• hK2
• Kallikrein 2
• kallikrein 2, prostatic
• kallikrein-2
• kallikrein-related peptidase 2
• Klk1c2
• KLK2
• KLK2A2
• MGC12201
• Tissue kallikrein-2
• Ton

Background

Kallikrein 2 (KLK2) is a secreted serine protease that is highly expressed in the human prostate gland (1). It is also known as the prostate-specific glandular kallikrein. The enzyme is highly specific for cleavage after arginine residues. KLK2 is able to activate the urokinase-type plasminogen activator (2). KLK2 is inhibited by serpins such as protein C inhibitor, antichymotrypsin, and plasminogen activator inhibitor 1 (3 ‑ 5). KLK2 is structurally related to KLK3, the prostate-specific antigen (PSA). Like PSA, KLK2 is considered to be a biomarker for prostate cancer (6).

1. Chapelaine, P. et al. (1988) FEBS Lett. 236:205.
2. Frenette, G. et al. (1997) Int. J. Cancer 71:897.
3. Deperthes, D. et al. (1995) Biochim. Biophys. Acta 1245:311.
4. Grauer, L.S. et al. (1998) J. Androl. 19:407.
5. Mikolajczyk S.D. et al. (1999) Cancer Res. 59:3927.
6. Sardana, G. et al. (2008) Clin. Chem. 54:1951.

Bioinformatics

• Gene Symbol: KLK2
• Uniprot:
  • P20151()