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Recombinant Human Kallikrein 5 Protein, CF

Catalog #: 1108-SE
19 Citations 1 Review Datasheet
Catalog # Availability Size / Price Qty
1108-SE-010
Recombinant Human Kallikrein 5 Protein Enzyme Activity
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Product Details
Citations (19)
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Reviews (1)
Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Kallikrein 5 Protein, CF Summary

Product Specifications

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 #
Q9Y337
N-terminal Sequence
Analysis
Ile67
Structure / Form
Mature form
Predicted Molecular Mass
27 kDa
SDS-PAGE
37-42 kDa, reducing conditions

Product Datasheets

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1108-SE

Product Datasheet

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.

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 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A9891)

Per Well:
  • rhKLK5: 0.1 µg
  • Substrate: 100 µM


























Scientific Data

Enzyme Activity Recombinant Human Kallikrein 5 Protein Enzyme Activity View Larger

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

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.

19 Citations: Showing 1 - 10
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  1. Betula alba Bark Extract and Empetrum nigrum Fruit Juice, a Natural Alternative to Niacinamide for Skin Barrier Benefits
    Authors: S Smiljanic, C Messaraa, V Lafon-Kolb, N Hrapovic, N Amini, C Osterlund, L Visdal-Joh
    International Journal of Molecular Sciences, 2022-10-19;23(20):.
    Species: Plant - Betula alba, Plant - Empetrum nigrum
    Sample Types: Complex Sample Type
    Applications: Bioassay
  2. Chlorogenic Acid Isomers Isolated from Artemisia lavandulaefolia Exhibit Anti-Rosacea Effects In Vitro
    Authors: KB Roh, Y Jang, E Cho, D Park, DH Kweon, E Jung
    Biomedicines, 2022-02-16;10(2):.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay
  3. 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, 2017-10-27;22(11):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  4. 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, 2017-09-27;6(11):1529-1539.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  5. 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., 2017-07-27;0(0):.
    Species: Virus
    Sample Types: Protein
    Applications: Enzyme Assay
  6. Protease-inhibitor interaction predictions: Lessons on the complexity of protein-protein interactions
    Authors: N Fortelny, GS Butler, CM Overall, P Pavlidis
    Mol. Cell Proteomics, 2017-04-06;0(0):.
    Applications: Bioassay
  7. 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, 2017-03-30;12(3):e0174827.
    Species: Virus - Influenza A
    Sample Types: Peptide
    Applications: Enzyme Assay
  8. Anti-inflammatory and immunomodulatory effects of Aquaphilus dolomiae extract on in vitro models
    Authors: Nathalie Castex-Riz
    Clin Cosmet Investig Dermatol, 2016-11-10;9(0):421-434.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  9. Defects in Stratum Corneum Desquamation Are the Predominant Effect of Impaired ABCA12 Function in a Novel Mouse Model of Harlequin Ichthyosis
    PLoS ONE, 2016-08-23;11(8):e0161465.
    Species: Human, Mouse
    Sample Types: In Vivo, Whole Cells
    Applications: Bioassay, In Vivo
  10. 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, 2014-05-28;289(29):20026-38.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  11. 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, 2013-01-10;133(5):1170-7.
    Applications: Bioassay
  12. 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, 2011-06-23;131(11):2223-32.
    Species: Human
    Sample Types: Substrate
    Applications: Enzyme Assay
  13. 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, 2011-03-23;32(6):1187-92.
    Applications: Bioassay
  14. 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., 2010-07-28;285(42):32174-81.
    Species: Human
    Sample Types: Peptide
    Applications: Enzyme Assay
  15. 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., 2010-07-25;42(8):676-83.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  16. 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., 2010-01-21;130(5):1297-306.
    Applications: ELISA (Standard)
  17. 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., 2009-08-13;422(2):383-92.
    Species: Human
    Sample Types: Antibody
    Applications: Surface Plasmon Resonance
  18. 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., 2008-01-25;275(5):1003-17.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  19. 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., 2007-08-05;13(8):975-80.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo

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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