>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<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 >150 pmol/min/µg, as measured under the described conditions. See Activity Assay Protocol on www.RnDSystems.com.
Mouse myeloma cell line, NS0-derived Glu23-His252, with a C-terminal 10-His tag
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
Dilute rhKLK7 to 200 μg/mL in Activation Buffer.
Dilute Thermolysin to 20 μg/mL in Activation Buffer.
Combine equal volumes of 200 μg/mL rhKLK7 and 20 μg/mL Thermolysin.
Incubate at 37 °C for 2 hours.
Stop reaction with an equal volume 100 mM EDTA diluted in Assay Buffer (Cf = 50 mM).
Dilute activated rhKLK7 to 4 ng/μL in Assay Buffer.
Dilute Substrate to 20 μM in Assay Buffer.
Load 50 μL of 4 ng/μL rhKLK7 in a plate, and start the reaction by adding 50 μL of 20 μM Substrate to each well. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 20 μM Substrate.
Read at excitation and emission wavelengths of 320 nm and 405 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
rhKLK7: 0.2 μg
Substrate: 10 μM
Background: Kallikrein 7
Human tissue Kallikrein 7 (hK7), also known as stratum corneum chymotryptic enzyme (SCCE), is a member of the human tissue kallikrein family. Full-length hK7 consists of 253 amino acids, with a signal peptide (residues 1‑22), short pro peptide (residues 23‑29) and mature chain (residues 30‑252) (1). Predominantly expressed in the skin, a major physiological function of hK7 is to regulate the desquamation process through proteolysis of the intercellular adhesive structures between corneocytes (2). Thus, it is related to some inflammatory skin diseases, such as psoriasis and chronic itchy dermatitis (3, 4). Studies have shown that one potential physiological activator for hK7 is hK5, another member of the human tissue Kallikrein family. Along with hK14, these three kallikreins form a proteolytic cascade in the stratum corneum (5). The purified, secreted recombinant human K7 corresponds to the pro form. When activated by thermolysin, it displays enzymatic activity towards a fluorogenic synthetic peptide described in the Activity Assay Protocol. This activity can be inhibited by recombinant human Serpin A1, A3, A4, and A5 (Catalog # 1268-PI, 1295-PI, 1669-PI, and 1266-PI).
Hansson, L. et al. (1994) J. Bio. Chem. 269:19420.
Caubet, C. et al. (2004) J. Invest. Dermatol. 122:1235.
Ekholm, E. and Egelrud, T. (1999) Arch. Dermatol. Res. 291:195.
Hansson, L. et al. (2002) J. Invest. Dermatol. 118:444.
Brattsand, M. et al. (2004) J. Invest. Dermatol. 124:198.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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