Recombinant Human Kallikrein 7 Protein, CF Summary
Glu23-His252, with a C-terminal 10-His tag
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.
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.
|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.
- 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
- 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).Per Well:
- 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.
Citations for Recombinant Human Kallikrein 7 Protein, CF
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.
Citations: Showing 1 - 9
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Loss of kallikrein-related peptidase 7 exacerbates amyloid pathology in Alzheimer's disease model mice
Authors: K Kidana, T Tatebe, K Ito, N Hara, A Kakita, T Saito, S Takatori, Y Ouchi, T Ikeuchi, M Makino, TC Saido, M Akishita, T Iwatsubo, Y Hori, T Tomita
EMBO Mol Med, 2018;0(0):.
Sample Types: Whole Cells
Protease-inhibitor interaction predictions: Lessons on the complexity of protein-protein interactions
Authors: N Fortelny, GS Butler, CM Overall, P Pavlidis
Mol. Cell Proteomics, 2017;0(0):.
Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation.
Authors: Yamamoto-Tanaka M, Motoyama A, Miyai M, Matsunaga Y, Matsuda J, Tsuboi R, Hibino T
J Biol Chem, 2014;289(29):20026-38.
Sample Types: Whole Cells
Proteolytic activation cascade of the Netherton syndrome-defective protein, LEKTI, in the epidermis: implications for skin homeostasis.
Authors: Fortugno P, Bresciani A, Paolini C, Pazzagli C, El Hachem M, D'Alessio M, Zambruno G
J Invest Dermatol, 2011;131(11):2223-32.
Sample Types: Substrate
Applications: Enzyme Assay
Matriptase initiates activation of epidermal pro-kallikrein and disease onset in a mouse model of Netherton syndrome.
Authors: Sales KU, Masedunskas A, Bey AL
Nat. Genet., 2010;42(8):676-83.
Sample Types: Recombinant Protein
Kallikrein expression and cathelicidin processing are independently controlled in keratinocytes by calcium, vitamin D(3), and retinoic acid.
Authors: Morizane S, Yamasaki K, Kabigting FD, Gallo RL
J. Invest. Dermatol., 2010;130(5):1297-306.
Sample Types: N/A
Applications: ELISA (Standard)
Isolation of SPINK6 in human skin: selective inhibitor of kallikrein-related peptidases.
Authors: Meyer-Hoffert U, Wu Z, Kantyka T, Fischer J, Latendorf T, Hansmann B, Bartels J, He Y, Glaser R, Schroder JM
J. Biol. Chem., 2010;285(42):32174-81.
Sample Types: Peptide
Applications: Enzyme Assay
Specific inhibition of tissue kallikrein 1 with a human monoclonal antibody reveals a potential role in airway diseases.
Authors: Sexton DJ, Chen T, Martik D, Kuzmic P, Kuang G, Chen J, Nixon AE, Zuraw BL, Forteza RM, Abraham WM, Wood CR
Biochem. J., 2009;422(2):383-92.
Sample Types: Antibody
Applications: Surface Plasmon Resonance
Inhibition of kallikrein-related peptidases by the serine protease inhibitor of Kazal-type 6.
Authors: Kantyka T, Fischer J, Wu Z, Declercq W, Reiss K, Schroder J, Meyer-Hoffert U
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Fluorogenic Peptide Substrates
Reviews for Recombinant Human Kallikrein 7 Protein, CF
Average Rating: 5 (Based on 3 Reviews)
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Reason for Rating: Enzymatic activity and positive controls for in vitro studies