Kallikrein 4 (KLK4), also known as prostase or enamel matrix serine protease 1 (EMSP1), is a serine protease of the human tissue kallikrein gene family (1). Among normal tissues, human KLK4 is specifically expressed in the prostate (2). It is over-expressed in prostate cancer and this expression is regulated by hormones including androgens, esterogen and progesterone (3). rhKLK4 readily activates pro-KLK3/PSA and pro-urokinase type plasminogen activator (uPA), indicating it may initiate events involving PSA and uPA in either normal or abnormal processes (4). KLK4 may have additional roles such as functioning as one of the two major enamel proteases identified that process enamel matrix proteins (5). In addition to being a secreted enzyme, it is also a nuclear protein (3, 6). The deduced amino acid sequence of human KLK4 consists of a signal peptide, a short pro region and a mature/active enzyme. rhKLK4 can be activated in vitro by thermolysin, a zinc protease. The peptidase activity can be inhibited by AEBSF (R&D Systems, Catalog # EI001), a general serine protease inhibitor.
Recombinant Human Kallikrein 4/Prostase Protein, CF
R&D Systems | Catalog # 1719-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Kallikrein 4/Prostase Protein (1719-SE)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Structure / Form
Pro form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 4/Prostase/EMSP1 protein
Ser27-Ser254, with a C-terminal 10-His tag
Ser27-Ser254, with a C-terminal 10-His tag
Purity
>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.
N-terminal Sequence Analysis
Ser27 & Ser29
Predicted Molecular Mass
26 kDa
SDS-PAGE
33 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).
The specific activity is >250 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 1719-SE in the following applications:
Formulation, Preparation, and Storage
1719-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile 50 mM Tris, 10 mM CaCl2 and 150 mM NaCl, pH 7.5. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Calculators
Background: Kallikrein 4/Prostase/EMSP1
References
- Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
- Nelson, P.S. et al. (1999) Proc. Natl. Acad. Sci. USA 96:3114.
- Xi, Z. et al. (2004) Cancer Res. 64:2365.
- Takayama, T.K. et al. (2001) Biochemistry 40:15341.
- Simmer, J.P. and J.C. Hu (2002) Connect Tissue Res. 43:441.
- Ryu, O.H. et al. (2002) Eur. J. Oral. Sci. 110:358.
Alternate Names
ARM1, EMSP1, KLK4, Prostase
Entrez Gene IDs
9622 (Human)
Gene Symbol
KLK4
UniProt
Additional Kallikrein 4/Prostase/EMSP1 Products
Product Documents for Recombinant Human Kallikrein 4/Prostase Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Kallikrein 4/Prostase Protein, CF
For research use only
Citations for Recombinant Human Kallikrein 4/Prostase Protein, CF
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Protocols
View specific protocols for Recombinant Human Kallikrein 4/Prostase Protein, CF (1719-SE):
Materials
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
- Assay Buffer: 50 mM Tris, pH 9.0
- Recombinant Human Kallikrein 4/Prostase/EMSP1 (rhKLK4) (Catalog # 1719-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
- Substrate: BOC-Val-Pro-Arg-AMC (Catalog # ES011), 90 mM Stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhKLK4 to 200 µg/mL in Activation Buffer.
- Dilute Thermolysin to 2 µg/mL in Activation Buffer.
- Combine equal volumes diluted rhKLK4 and Thermolysin and incubate at 37 °C for 2 hours to activate.
- Stop the reaction with 1,10 Phenanthroline at a final concentration of 10 mM.
- Dilute activated rhKLK4 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 µL of the 2 ng/μL rhKLK4 in a plate and start the reaction by adding 50 μL of 200 μM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate without any rhKLK4.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 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).
Per Well:
- rhKLK4: 0.100 µg
- Substrate: 100 µM