Recombinant Human Kallikrein 5 Protein, CF

Catalog # Availability Size / Price Qty
1108-SE-010
Kallikrein 5 Protein Enzyme Activity
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Product Details
Citations (17)
FAQs
Supplemental Products
Reviews (1)

Recombinant Human Kallikrein 5 Protein, CF Summary

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
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 Boc-VPR-AMC (Catalog # ES011). The specific activity is >200 pmol/min/µg, as measured under the described conditions. 
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 5 protein
Ile67-Ser293, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Ile67
Structure / Form
Mature form
Predicted Molecular Mass
31 kDa
SDS-PAGE
40 kDa, reducing conditions

Product Datasheets

1108-SE

Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and Glycerol.
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
  • Assay Buffer: 0.1 M NaH2PO4, pH 8.0
  • Recombinant Human Kallikrein 5 (rhKLK5) (Catalog # 1108-SE)
  • Fluorogenic Peptide Substrate BOC-Val-Pro-Arg-AMC (Catalog # ES011), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhKLK5 to 2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load 50 µL of the 2 ng/µL rhKLK5 into plate and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate without any rhKLK5.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. 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:
  • rhKLK5: 0.1 µg
  • Substrate: 100 µM

Data Image

Enzyme Activity Kallikrein 5 Protein Enzyme Activity View Larger

Recombinant Human Kallikrein 5 (Catalog # 1108-SE) is measured byits ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).

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Background: Kallikrein 5

Human tissue Kallikrein 5 (hK5, also known as stratum corneum tryptic enzyme) is a serine protease of the human tissue Kallikrein gene family (2). The deduced amino acid sequence of human KLK5 consists of a signal peptide (Met1 to Gly22), a pro region (Val23 to Arg66) and a mature/active enzyme (Ile67 to Ser293). KLK5 is expressed mainly in skin, breast, brain and testis and to a lesser extent in many other tissues (3). KLK5 overexpression is an indicator of poor prognosis in ovarian cancer (4). hK5 is a candidate physiological activator of hK7 in the stratum corneum (5). Indeed, rhK5 can be used to activate rhK7 (R&D Systems, Catalog # 2624‑SE) by incubating rhK5 with recombinant human (rh) K7 (both at 0.1 mg/mL) in 25 mM MES, 0.15 M NaCl, pH 5.5, at 37 °C overnight.

rhK5 enzymatic activity can be inhibited by rhSerpinA1, A5, F2, and G1 (R&D Systems, Catalog # 1268‑PI, 1266‑PI, 1470-PI, 2488‑PI).

References
  1. Brattsand, M. and T. Egelrud (1999) J. Biol. Chem. 274:30033.
  2. Yousef, G.M. et al. (1999) Anticancer Res. 19:2843.
  3. Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
  4. Yousef, G.M. and E.P. Diamandis (1999) J. Biol. Chem. 274:37511.
  5. Kim, H. et al. (2001) Br. J. Cancer 84:643.
Entrez Gene IDs
25818 (Human); 68668 (Mouse); 690603 (Rat)
Alternate Names
EC 3.4.21; EC 3.4.21.4; Kallikrein 5; Kallikrein-like protein 2; kallikrein-related peptidase 5; KLK5; KLKL2; KLK-L2; KLK-L2EC 3.4.21.-; SCTE; SCTEkallikrein-5; Stratum corneum tryptic enzyme

Citations for Recombinant Human Kallikrein 5 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.

17 Citations: Showing 1 - 10
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  1. Kallikrein-related peptidase 5 contributes to H3N2 influenza virus infection in human lungs
    Authors: M Magnen, F Gueugnon, A Guillon, T Baranek, VC Thibault, A Petit-Cour, SJ de Veer, J Harris, AA Humbles, M Si-Tahar, Y Courty
    J. Virol., 2017;0(0):.
    Species: Virus
    Sample Types: Protein
    Applications: EnzAct
  2. KLK5 induces shedding of DPP4 from circulatory Th17 cells in type 2 diabetes
    Authors: T Nargis, K Kumar, AR Ghosh, A Sharma, D Rudra, D Sen, S Chakrabart, S Mukhopadhy, D Ganguly, P Chakrabart
    Mol Metab, 2017;6(11):1529-1539.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. A peptide-based approach to evaluate the adaptability of influenza A virus to humans based on its hemagglutinin proteolytic cleavage site
    Authors: MR Straus, GR Whittaker
    PLoS ONE, 2017;12(3):e0174827.
    Species: Influenza A Virus
    Sample Types: Peptide
    Applications: EnzAct
  4. 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
  5. Inhibition of Human Kallikrein 5 Protease by Triterpenoids from Natural Sources
    Authors: Y Matsubara, T Matsumoto, J Koseki, A Kaneko, S Aiba, K Yamasaki
    Molecules, 2017;22(11):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  6. Defects in Stratum Corneum Desquamation Are the Predominant Effect of Impaired ABCA12 Function in a Novel Mouse Model of Harlequin Ichthyosis
    PLoS ONE, 2016;11(8):e0161465.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. Anti-inflammatory and immunomodulatory effects of Aquaphilus dolomiae extract on in vitro models
    Authors: Nathalie Castex-Riz
    Clin Cosmet Investig Dermatol, 2016;9(0):421-434.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  8. 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
  9. Cross-linking of SPINK6 by transglutaminases protects from epidermal proteases.
    Authors: Fischer J, Koblyakova Y, Latendorf T, Wu Z, Meyer-Hoffert U
    J Invest Dermatol, 2013;133(5):1170-7.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay
  10. 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.
  11. 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
  12. 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
  13. 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
  14. 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
  15. Activation of hepatocyte growth factor activator zymogen (pro-HGFA) by human kallikrein 1-related peptidases.
    Authors: Mukai S, Fukushima T, Naka D, Tanaka H, Osada Y, Kataoka H
    FEBS J., 2008;275(5):1003-17.
    Species: Human
    Sample Types: Protein
    Applications: EnzAct
  16. Increased serine protease activity and cathelicidin promotes skin inflammation in rosacea.
    Authors: Yamasaki K, Di Nardo A, Bardan A, Murakami M, Ohtake T, Coda A, Dorschner RA, Bonnart C, Descargues P, Hovnanian A, Morhenn VB, Gallo RL
    Nat. Med., 2007;13(8):975-80.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  17. 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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Recombinant Human Kallikrein 5 Protein, CF
By Anonymous on 02/09/2017
Application: In vitro bioactivity in cell culture
Reason for Rating: we use it as standard in elisa