Recombinant Human Kallikrein 5 Protein, CF

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

Product Details

Summary
Reactivity HuSpecies Glossary
Applications EnzAct
Format
Carrier-Free

Order Details

Recombinant Human Kallikrein 5 Protein, CF Summary

Details of Functionality
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
Ile67-Ser293, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Ile67
Structure / Form
Mature form
Protein/Peptide Type
Recombinant Enzymes
Gene
KLK5
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
31 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
40 kDa, reducing conditions
Publications
Read Publications using
1108-SE in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in MES, NaCl and Glycerol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
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
  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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Kallikrein 5 Protein, CF

  • 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

Background

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).

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Publications for (11)

We have publications tested in 4 confirmed species: Human, Mouse, Influenza A Virus, N/A.

We have publications tested in 6 applications: Bioassay, ELISA Standard, EnzAct, Enzyme Assay, In Vivo, Surface Plasmon Resonance.


Filter By Application
Bioassay
(4)
ELISA Standard
(1)
EnzAct
(2)
Enzyme Assay
(1)
In Vivo
(1)
Surface Plasmon Resonance
(1)
All Applications
Filter By Species
Human
(6)
Mouse
(1)
Influenza A Virus
(1)
N/A
(2)
All Species
Showing Publications 1 - 10 of 11. Show All 11 Publications.
Publications using 1108-SE Applications Species
MR Straus, GR Whittaker A peptide-based approach to evaluate the adaptability of influenza A virus to humans based on its hemagglutinin proteolytic cleavage site PLoS ONE, 2017;12(3):e0174827. 2017 [PMID: 28358853] (EnzAct, Influenza A Virus) EnzAct Influenza A Virus
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. 2016 [PMID: 27551807] (Bioassay, Human) Bioassay Human
Yamamoto-Tanaka M, Motoyama A, Miyai M, Matsunaga Y, Matsuda J, Tsuboi R, Hibino T Mesotrypsin and caspase-14 participate in prosaposin processing: potential relevance to epidermal permeability barrier formation. J Biol Chem, 2014;289(29):20026-38. 2014 [PMID: 24872419] (Bioassay, Human) Bioassay Human
Fischer J, Koblyakova Y, Latendorf T, Wu Z, Meyer-Hoffert U Cross-linking of SPINK6 by transglutaminases protects from epidermal proteases. J Invest Dermatol, 2013;133(5):1170-7. 2013 [PMID: 23303447] (Bioassay, N/A) Bioassay N/A
Fortugno P, Bresciani A, Paolini C, Pazzagli C, El Hachem M, D'Alessio M, Zambruno G Proteolytic activation cascade of the Netherton syndrome-defective protein, LEKTI, in the epidermis: implications for skin homeostasis. J Invest Dermatol, 2011;131(11):2223-32. 2011 [PMID: 21697885]
Meyer-Hoffert U, Wu Z, Kantyka T, Fischer J, Latendorf T, Hansmann B, Bartels J, He Y, Glaser R, Schroder JM Isolation of SPINK6 in human skin: selective inhibitor of kallikrein-related peptidases. J. Biol. Chem., 2010;285(42):32174-81. 2010 [PMID: 20667819] (Enzyme Assay, Human) Enzyme Assay Human
Sales KU, Masedunskas A, Bey AL Matriptase initiates activation of epidermal pro-kallikrein and disease onset in a mouse model of Netherton syndrome. Nat. Genet., 2010;42(8):676-83. 2010 [PMID: 20657595] (Bioassay, Human) Bioassay Human
Morizane S, Yamasaki K, Kabigting FD, Gallo RL Kallikrein expression and cathelicidin processing are independently controlled in keratinocytes by calcium, vitamin D(3), and retinoic acid. J. Invest. Dermatol., 2010;130(5):1297-306. 2010 [PMID: 20090765] (ELISA Standard, N/A) ELISA Standard N/A
Sexton DJ, Chen T, Martik D, Kuzmic P, Kuang G, Chen J, Nixon AE, Zuraw BL, Forteza RM, Abraham WM, Wood CR Specific inhibition of tissue kallikrein 1 with a human monoclonal antibody reveals a potential role in airway diseases. Biochem. J., 2009;422(2):383-92. 2009 [PMID: 19527222] (Surface Plasmon Resonance, Human) Surface Plasmon Resonance Human
Mukai S, Fukushima T, Naka D, Tanaka H, Osada Y, Kataoka H Activation of hepatocyte growth factor activator zymogen (pro-HGFA) by human kallikrein 1-related peptidases. FEBS J., 2008;275(5):1003-17. 2008 [PMID: 18221492] (EnzAct, Human) EnzAct Human
Show All 11 Publications.

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Bioinformatics

Gene Symbol KLK5
Entrez
Uniprot