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Recombinant Human Kallikrein 7 Protein, CF

R&D Systems | Catalog # 2624-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Kallikrein 7 Protein (2624-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

P49862

Structure / Form

Pro form

Applications

Enzyme Activity
View all Kallikrein 7 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Kallikrein 7 protein
Glu23-His252, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Glu23

Predicted Molecular Mass

26 kDa

SDS-PAGE

29 kDa and 32 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 >150 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 3 reviews rated 5 using 2624-SE in the following applications:

Scientific Data Images for Recombinant Human Kallikrein 7 Protein, CF

Recombinant Human Kallikrein 7 Protein Enzyme Activity

Recombinant Human Kallikrein 7 Protein Enzyme Activity

Recombinant Human Kallikrein 7 (Catalog # 2624-SE) is measured by its ability to cleave the fluorogenic peptide substrate Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).

Formulation, Preparation, and Storage

2624-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

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 # http://www.rndsystems.com/product_results.aspx?k=1268-PI">1268-PI, http://www.rndsystems.com/product_results.aspx?k=1295-PI">1295-PI, http://www.rndsystems.com/product_results.aspx?k=1669-PI">1669-PI, and http://www.rndsystems.com/product_results.aspx?k=1266-PI">1266-PI).

References

  1. Hansson, L. et al. (1994) J. Bio. Chem. 269:19420.
  2. Caubet, C. et al. (2004) J. Invest. Dermatol. 122:1235.
  3. Ekholm, E. and Egelrud, T. (1999) Arch. Dermatol. Res. 291:195.
  4. Hansson, L. et al. (2002) J. Invest. Dermatol. 118:444.
  5. Brattsand, M. et al. (2004) J. Invest. Dermatol. 124:198.

Alternate Names

KLK7

Entrez Gene IDs

5650 (Human); 23993 (Mouse)

Gene Symbol

KLK7

UniProt

P49862

Additional Kallikrein 7 Products

Product Documents for Recombinant Human Kallikrein 7 Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Kallikrein 7 Protein, CF

For research use only

Citations for Recombinant Human Kallikrein 7 Protein, CF

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    Application: Enzymatic activity in vitro
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  • Recombinant Human Kallikrein 7 Protein, CF
    Name: Anonymous
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 05/02/2018
    Recombinant Human Kallikrein 7 Protein, CF 2624-SE
  • Recombinant Human Kallikrein 7 Protein, CF
    Name: Anonymous
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 08/10/2017

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Protocols

View specific protocols for Recombinant Human Kallikrein 7 Protein, CF (2624-SE):

Materials
  • Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Assay Buffer: 50 mM Tris, 150 mM NaCl, pH 8.5
  • Recombinant Human Kallikrein 7 (rhKLK7) (Catalog # 2624-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • EDTA (Sigma, Catalog # E-4884)
  • Substrate: MCA-Arg-Pro Lys-Pro-Val-Glu-Nval-Trp-Arg-Lys(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 rhKLK7 to 200 μg/mL in Activation Buffer.
  2. Dilute Thermolysin to 20 μg/mL in Activation Buffer.
  3. Combine equal volumes of 200 μg/mL rhKLK7 and 20 μg/mL Thermolysin.
  4. Incubate at 37 °C for 2 hours.
  5. Stop reaction with an equal volume 100 mM EDTA diluted in Assay Buffer (Cf = 50 mM).
  6. Dilute activated rhKLK7 to 4 ng/μL in Assay Buffer.
  7. Dilute Substrate to 20 μM in Assay Buffer.
  8. 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.
  9. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  10. 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:

  • rhKLK7: 0.2 μg
  • Substrate: 10 μM

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