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Recombinant Human Kallikrein 6 Protein, CF

Catalog #: 5164-SE
7 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
5164-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (7)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Kallikrein 6 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-QAR-AMC (Catalog # ES014). The specific activity is >1300 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 6/Neurosin protein
Glu17-Lys244, with a C-terminal 10-His tag
Accession #
Q92876
N-terminal Sequence
Analysis
Glu17
Structure / Form
Pro form
Predicted Molecular Mass
26 kDa
SDS-PAGE
35 kDa, reducing conditions

Product Datasheets

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5164-SE

Product Datasheet
COA
SDS

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.

5164-SE

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.
  • 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
  • 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
  1. Dilute rhKLK6 to 200 µg/mL in Activation Buffer.
  2. Dilute Lysyl-endopeptidase to 2.5 mU/mL in Activation Buffer.
  3. Combine and mix equal volumes of diluted rhKLK6 and diluted Lysyl-endopeptidase.
  4. Incubate at room temperature for 30 minutes.
  5. After incubation, dilute rhKLK6 to 0.5 µg/mL in Assay Buffer.
  6. Dilute Substrate to 200 µM in Assay Buffer.
  7. 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.
  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:
  • rhKLK6: 0.025 µg
  • Substrate: 100 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

References
  1. Emami, N. and E.P. Diamandis (2007) Clin Chim Acta 381:78.
  2. Borgoño, C.A. and E.P. Diamandis (2004) Nat. Rev. Cancer, 4:876.
  3. Paliouras, M. et al. (2007) Cancer Lett. 249:61.
Entrez Gene IDs
5653 (Human)
Alternate Names
Bssp; EC 3.4.21; EC 3.4.21.-; hK6; kallikrein 6 (neurosin, zyme); Kallikrein 6; kallikrein-6; kallikrein-related peptidase 6; KLK6; Klk7; Neurosin; Protease M; protease, serine, 18; PRSS18; PRSS9; PRSS9MGC9355; Serine protease 18; Serine protease 9; SP59; Zyme

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.

7 Citations: Showing 1 - 7
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  1. Genuine selective caspase-2 inhibition with new irreversible small peptidomimetics
    Authors: E Bosc, J Anastasie, F Soualmia, P Coric, JY Kim, LQ Wang, G Lacin, K Zhao, R Patel, E Duplus, P Tixador, AA Sproul, B Brugg, M Reboud-Rav, CM Troy, ML Shelanski, S Bouaziz, M Karin, C El Amri, ED Jacotot
    Cell Death & Disease, 2022-11-15;13(11):959.
    Species: N/A
    Sample Types: Fluorogenic Peptide Substrate
    Applications: Bioassay
  2. 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
    Oncogene, 2022-07-19;12(7):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. 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, 2018-03-01;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. 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, 2013-01-08;8(1):e53250.
    Applications: ELISA (Standard), Western Blot
  5. 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, 2012-04-17;21(1):31-41.
    Applications: Bioassay
  6. 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
  7. 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

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