Recombinant Human Kallikrein 7 Protein, CF

Catalog # Availability Size / Price Qty
2624-SE-010
Recombinant Human Kallikrein 7 Protein Enzyme Activity
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Product Details
Citations (9)
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Reviews (3)

Recombinant Human Kallikrein 7 Protein, CF Summary

Product Specifications

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.
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.
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 7 protein
Glu23-His252, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Glu23
Structure / Form
Pro form
Predicted Molecular Mass
26 kDa
SDS-PAGE
29 kDa and 32 kDa, reducing conditions

Product Datasheets

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.

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.

Assay Procedure

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

Data Image

Enzyme Activity Recombinant Human Kallikrein 7 Protein Enzyme Activity View Larger

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).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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-PI1669-PI, and 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.
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

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.

9 Citations: Showing 1 - 9
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  1. 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):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. 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):.
    Applications: Bioassay
  3. 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.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. 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.
    Species: Human
    Sample Types: Substrate
    Applications: Enzyme Assay
  5. 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.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  6. 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.
    Species: N/A
    Sample Types: N/A
    Applications: ELISA (Standard)
  7. 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.
    Species: Human
    Sample Types: Peptide
    Applications: Enzyme Assay
  8. 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.
    Species: Human
    Sample Types: Antibody
    Applications: Surface Plasmon Resonance
  9. 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
    Peptides, 0;32(6):1187-92.
    Applications: Bioassay

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Reviews for Recombinant Human Kallikrein 7 Protein, CF

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Recombinant Human Kallikrein 7 Protein, CF
By Anonymous on 03/13/2020
Application: Enzymatic activity in vitro

Recombinant Human Kallikrein 7 Protein, CF
By Anonymous on 05/02/2018
Application: Enzymatic activity in vitro
Reason for Rating: Enzymatic activity and positive controls for in vitro studies

Recombinant Human Kallikrein 7 Protein, CF
By Anonymous on 08/10/2017
Application: Enzymatic activity in vitro