Recombinant Mouse Kallikrein 7 Protein, CF

R&D Systems | Catalog # 7688-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Kallikrein 7 Protein (7688-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

Q91VE3

Applications

Enzyme Activity
View all Kallikrein 7 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Kallikrein 7 protein
Gln22-Arg249 with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

No results obtained: Gln22 predicted

Predicted Molecular Mass

26 kDa

SDS-PAGE

27-37 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >70 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 7688-SE in the following applications:

Formulation, Preparation, and Storage

7688-SE
Formulation Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
Shipping The product is shipped with polar packs. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Kallikrein 7

Kallikrein 7 (KLK7), also known as stratum corneum chymotryptic enzyme (SCCE), is a member of the tissue kallikrein family.  Predominantly expressed in the skin, a major physiological function of KLK7 is to regulate the desquamation process through proteolysis of the intercellular adhesive structures between corneocytes (1). Thus, it is involved in some inflammatory skin diseases, such as psoriasis and chronic itchy dermatitis (2, 3). Studies have shown that one potential physiological activator for KLK7 is KLK5, another member of the tissue kallikrein family. Along with KLK14, these three kallikreins form a proteolytic cascade in the stratum corneum (4). Mouse KLK7 is synthesized as a 249 amino acid precursor with a 21 amino acid signal peptide. The secreted protein has a short propeptide (residues 22‑25) and a mature chain comprised of residues 26‑249. The purified, secreted rmKLK7 corresponds to the pro form. After activation by thermolysin, it displays enzymatic activity against a fluorogenic synthetic peptide as described in the Activity Assay Protocol.

References

  1. Caubet, C. et al. (2004) J. Invest. Dermatol. 122:1235.
  2. Ekholm, E. and Egelrud, T. (1999) Arch. Dermatol. Res. 291:195.
  3. Hansson, L. et al. (2002) J. Invest. Dermatol. 118:444.
  4. Brattsand, M. et al. (2004) J. Invest. Dermatol. 124:198.

Alternate Names

KLK7

Entrez Gene IDs

5650 (Human); 23993 (Mouse)

Gene Symbol

KLK7

UniProt

Q91VE3 (Mouse)

Additional Kallikrein 7 Products

Product Documents for Recombinant Mouse Kallikrein 7 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.

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Product Specific Notices for Recombinant Mouse Kallikrein 7 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

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Protocols

View specific protocols for Recombinant Mouse Kallikrein 7 Protein, CF (7688-SE):

Materials
  • Activation Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% Brij-35, pH 7.5 (TCNB)
  • Assay Buffer: 50 mM Tris, 1 M NaCl, pH 8.0
  • Recombinant Mouse Kallikrein 7 (rmKLK7) (Catalog # 7688-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • EDTA, Disodium salt (Sigma, Catalog # E-4884) 500 mM stock in deionized water
  • Substrate: Mca-RPKPVE-NVal-WRK(Dnp)-NH2 (Catalog # ES002)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmKLK7 to 200 µg/mL in Activation Buffer.
  2. Dilute Thermolysin to 20 µg/mL in Activation Buffer.
  3. Activate rmKLK7 by combining equal volumes of 200 µg/mL rmKLK7 with 20 µg/mL Thermolysin.
  4. Incubate reactions at 37 °C for two hours.
  5. Stop reactions by adding EDTA to a final concentration of 50 mM.
  6. Dilute activated rmKLK7 to 2 µg/mL in Assay Buffer.
  7. Dilute Substrate to 20 µM in Assay Buffer.
  8. In a plate, load 50 µL of activated rmKLK7. Include an enzyme blank containing 50 µL Assay Buffer.
  9. Start reaction by adding 50 µL of 20 µM 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:

  • rmKLK7: 0.1 µg
  • Substrate: 10 µM

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