Recombinant Human Kallikrein 5 Protein, CF Summary
Product Specifications
Ile67-Ser293, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
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Assay Procedure
- 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
- Dilute rhKLK5 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 380 nm and 460 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 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A9891)
Per Well:- rhKLK5: 0.1 µg
- Substrate: 100 µM
Scientific Data
Recombinant Human Kallikrein 5 (Catalog # 1108-SE) is measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).
Reconstitution Calculator
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).
- Brattsand, M. and T. Egelrud (1999) J. Biol. Chem. 274:30033.
- Yousef, G.M. et al. (1999) Anticancer Res. 19:2843.
- Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
- Yousef, G.M. and E.P. Diamandis (1999) J. Biol. Chem. 274:37511.
- Kim, H. et al. (2001) Br. J. Cancer 84:643.
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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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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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 -
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) -
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 -
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 -
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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