Recombinant Human Kallikrein 6 Protein, CF Summary
Glu17-Lys244, with a C-terminal 10-His tag
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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.
|Formulation||Supplied as a 0.2 μm filtered solution in Sodium Citrate.|
|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.
- Activation Buffer: 50 mM Tris, 0.05% (w/v) Brij-35, pH 8.0
- Assay Buffer: 50 mM Tris, 1.0 M Sodium Citrate, pH 7.5
- Recombinant Human Kallikrein 6/Neurosin (rhKLK6) (Catalog # 5164-SE)
- Lysyl-Endopeptidase (Wako BioProducts, Catalog # 129-02541), 20 U/mL in PBS
- Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014), 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 rhKLK6 to 200 µg/mL in Activation Buffer.
- Dilute Lysyl-endopeptidase to 2.5 mU/mL in Activation Buffer.
- Combine and mix equal volumes of diluted rhKLK6 and diluted Lysyl-endopeptidase.
- Incubate at room temperature for 30 minutes.
- After incubation, dilute rhKLK6 to 0.5 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load into a black well plate 50 µL of 0.5 µg/mL rhKLK6, 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.
- 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 # A-9891).Per Well:
- rhKLK6: 0.025 µg
- Substrate: 100 µM
Background: Kallikrein 6/Neurosin
Tissue kallikreins are a family of extracellular serine proteases consisting of 15 members (1, 2). The genes of this family aligned in tandem on chromosome 19q13.4 represent the largest contiguous group of proteases within the human genome. Tissue kallikreins have attracted great interest as potential biomarkers for various cancers, including prostate, ovarian, breast, testicular, and lung (2, 3). Human Kallikrein 6 (hKLK6) is a member of tissue kallikrein family observed in breast and brain tissues, colon carcinoma cells, and oligodedrocytes (1 - 3). Known protein substrates of hKLK6 are myelin basic protein, the precursor of the A beta amyloid peptide, and plasminogen. Its physiological functions may include the participation in demyelination processes as well as in the progression of inflammatory disease of the CNS. The level of hKLK6 is reduced in brain extracts of Alzheimer patients and increased in serum of patients with ovarian cancer.
- Emami, N. and E.P. Diamandis (2007) Clin Chim Acta 381:78.
- Borgoño, C.A. and E.P. Diamandis (2004) Nat. Rev. Cancer, 4:876.
- Paliouras, M. et al. (2007) Cancer Lett. 249:61.
Citations for Recombinant Human Kallikrein 6 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.
Citations: Showing 1 - 6
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Kallikrein-Related Peptidase 6 (KLK6) as a Contributor toward an Aggressive Cancer Cell Phenotype: A Potential Role in Colon Cancer Peritoneal Metastasis
Authors: H Bouzid, F Soualmia, K Oikonomopo, A Soosaipill, F Walker, K Louati, R Lo Dico, M Pocard, C El Amri, NA Ignatenko, D Darmoul
Sample Types: Whole Cells
Loss of kallikrein-related peptidase 7 exacerbates amyloid pathology in Alzheimer's disease model mice
Authors: K Kidana, T Tatebe, K Ito, N Hara, A Kakita, T Saito, S Takatori, Y Ouchi, T Ikeuchi, M Makino, TC Saido, M Akishita, T Iwatsubo, Y Hori, T Tomita
EMBO Mol Med, 2018;0(0):.
Sample Types: Whole Cells
Low CSF Levels of Both alpha-Synuclein and the alpha-Synuclein Cleaving Enzyme Neurosin in Patients with Synucleinopathy.
Authors: Wennstrom M, Surova Y, Hall S, Nilsson C, Minthon L, Bostrom F, Hansson O, Nielsen H
PLoS ONE, 2013;8(1):e53250.
Sample Types: N/A
Applications: ELISA (Standard)
Lentivirus mediated delivery of neurosin promotes clearance of wild-type alpha-synuclein and reduces the pathology in an alpha-synuclein model of LBD.
Authors: Spencer B, Michael S, Shen J, Kosberg K, Rockenstein E, Patrick C, Adame A, Masliah E
Mol Ther, 2013;21(1):31-41.
Sample Types: Recombinant Protein
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.
Sample Types: Antibody
Applications: Surface Plasmon Resonance
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
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Fluorogenic Peptide Substrates
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