Recombinant Human Kallikrein 14 Protein, CF

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Recombinant Human Kallikrein 14 (Catalog # 2626‑SE) ismeasured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Kallikrein 14 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 >3,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 14 protein
Gln19-Met248, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
No results obtained: Gln19 predicted
Structure / Form
Pro form
Protein/Peptide Type
Recombinant Enzymes
Gene
KLK14
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
26 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
28 kDa, reducing conditions
Publications
Read Publications using
2626-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 Tris, NaCl and Glycerol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Assay Procedure
  • 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, 0.05% (w/v) Brij-35, pH 8.0
  • Recombinant Human Kallikrein 14 (rhKLK14) (Catalog # 2626-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • EDTA  (Sigma, Catalog # E-4884)
  • Substrate:  BOC-Val-Pro-Arg-AMC (Catalog # ES011), 100 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate reader (Model: SpectramMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhKLK14 to 200 µg/mL in Activation Buffer.
  2. Dilute Thermolysin to 20 µg/mL in Activation Buffer.
  3. Combine 25 µL of diluted rhKLK14 and 25 µL of diluted Thermolysin.
  4. Incubate at 37 °C for 1 hour.
  5. Stop reaction with 50 µL of 100 mM EDTA.
    Dilute incubated rhKLK14 mixture to 0.2 ng/µL in Assay Buffer.
  6. Dilute Substrate to 200 µM in Assay Buffer.
  7. Load 50 µL of the 0.2 ng/µL rhKLK14 mixture in a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
  8. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively in kinetic mode for 5 minutes.
  9. 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 # A-9891).

Per Well:
  • rhKLK14: 0.01 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 14 Protein, CF

  • EC 3.4.21
  • EC 3.4.21.-
  • hK14
  • Kallikrein 14
  • Kallikrein-like protein 6
  • kallikrein-related peptidase 14
  • KLK14
  • KLKL6
  • KLK-L6kallikrein-14

Background

Human tissue kallikreins refer to a group of secreted serine proteases that are encoded by homologous genes clustering on chromosome 19q13.3-4. As a member of this family, human tissue Kallikrein 14 (hKLK14) is present in many tissues, with high levels in breast, skin, prostate, and brain. The 251 amino acid hKLK14 precursor consists of a signal peptide (residues 1 to 18), a pro peptide (residues 19 to 24) and an active protein (residues 25 to 251) (1). Its enzymatic activity has been shown to be mainly trypsin-like (2). However, its physiological substrates and functions are still unclear. Several studies have suggested that hKLK14 may have clinical utility as a biomarker for cancer of the breast, ovary, and prostate (3, 4). In addition, it may be the initiator of a kallikrein proteolytic cascade responsible for the degradation of the adhesion structures in the stratum corneum (2). The purified, secreted rhKLK14 corresponds to the pro form (residues 19 to 248) with a replacement of the last three residues with a His tag at the C-terminus. When activated by thermolysin, it displays enzymatic activity towards a fluorogenic peptide described above. This activity can be inhibited by recombinant human Serpin A4, E1, and F2 (Catalog # 1669-PI1786-PI, and 1470-PI, respectively).

  1. Yousef, G. M. et al. (2001) Cancer Res. 61:3425.
  2. Brattsand, M. et al. (2004) J. Invest. Dermatol. 124:198.
  3. Borgono, C. et al. (2003) Cancer Res. 63:9032.
  4. Yousef, G. M. et al. (2003) Prostate. 56:287.

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2626-SE
Species: Hu
Applications: Enzyme Activity

Publications for Kallikrein 14 (2626-SE)(6)

We have publications tested in 2 confirmed species: Human, N/A.

We have publications tested in 4 applications: Bioassay, Enzyme Activity, Enzyme Assay, Surface Plasmon Resonance.


Filter By Application
Bioassay
(2)
Enzyme Activity
(2)
Enzyme Assay
(1)
Surface Plasmon Resonance
(1)
All Applications
Filter By Species
Human
(4)
N/A
(1)
All Species
Showing Publications 1 - 6 of 6.
Publications using 2626-SE Applications Species
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, Human) Bioassay Human
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] (Enzyme Activity, Human) Enzyme Activity Human
Kantyka T, Fischer J, Wu Z, Declercq W, Reiss K, Schroder J, Meyer-Hoffert U Inhibition of kallikrein-related peptidases by the serine protease inhibitor of Kazal-type 6. Peptides, 0;32(6):1187-92. 0 [PMID: 21439340] (Bioassay) Bioassay
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
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
Yoon H, Laxmikanthan G, Lee J, Blaber SI, Rodriguez A, Kogot JM, Scarisbrick IA, Blaber M Activation profiles and regulatory cascades of the human kallikrein-related peptidases. J. Biol. Chem., 2007;282(44):31852-64. 2007 [PMID: 17823117] (Enzyme Activity, N/A) Enzyme Activity N/A

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Bioinformatics

Gene Symbol KLK14
Uniprot