Recombinant Human Kallikrein 2 Protein, CF Summary
Product Specifications
Met1-Pro261, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
4104-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Sodium Citrate 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.
|
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
- Dilute rhKLK2 to 4 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 0 to 5 minutes.
- 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).
- rhKLK2: 0.2 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: Kallikrein 2
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).
- Chapelaine, P. et al. (1988) FEBS Lett. 236:205.
- Frenette, G. et al. (1997) Int. J. Cancer 71:897.
- Deperthes, D. et al. (1995) Biochim. Biophys. Acta 1245:311.
- Grauer, L.S. et al. (1998) J. Androl. 19:407.
- Sardana, G. et al. (2008) Clin. Chem. 54:1951.
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