Recombinant Mouse Kallikrein 7 Protein, CF

Name: Recombinant Mouse Kallikrein 7 Protein, CF
Brand: R&D Systems
SKU: 7688-SE
Rating: 5 (1 reviews)

Catalog #: 7688-SE 1 Review Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
7688-SE-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Kallikrein 7 Protein, CF Summary

Product Specifications

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.

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.

Source

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

Accession #

Q91VE3

N-terminal Sequence
Analysis

No results obtained: Gln22 predicted

Predicted Molecular Mass

26 kDa

SDS-PAGE

27-37 kDa, reducing conditions

Product Datasheets

7688-SE

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

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.

Assay Procedure

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

Dilute rmKLK7 to 200 µg/mL in Activation Buffer.
Dilute Thermolysin to 20 µg/mL in Activation Buffer.
Activate rmKLK7 by combining equal volumes of 200 µg/mL rmKLK7 with 20 µg/mL Thermolysin.
Incubate reactions at 37 °C for two hours.
Stop reactions by adding EDTA to a final concentration of 50 mM.
Dilute activated rmKLK7 to 2 µg/mL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
In a plate, load 50 µL of activated rmKLK7. Include an enzyme blank containing 50 µL Assay Buffer.
Start reaction by adding 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 V_max* (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

Reconstitution Calculator

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

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.

Entrez Gene IDs

5650 (Human); 23993 (Mouse)

Alternate Names

EC 3.4.21; EC 3.4.21.117; hK7; hSCCE; kallikrein 7 (chymotryptic, stratum corneum); Kallikrein 7; kallikrein-related peptidase 7; KLK7; protease, serine, 6; PRSS6; SCCEkallikrein-7; Serine protease 6; signal protein; Stratum corneum chymotryptic enzyme