Recombinant Human Kallikrein 13 Protein, CF Summary
Product Specifications
Met1-Ile262, with a C-terminal 6-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.
2625-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.
|
Assay Procedure
- Activation Buffer: 0.1 M Tris, pH 8.0
- Assay Buffer: 50 mM Tris, 150 mM NaCl, 0.05% Brij-35 (v/v), pH 8.0
- Recombinant Human Kallikrein 13 (rhKLK13) (Catalog # 2625-SE)
- Lysyl-Endopeptidase (Wako BioProducts, Catalog # 129-02541)
- Substrate: Boc-Val-Pro-Arg-AMC (Catalog # ES011), 100 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhKLK13 to 100 µg/mL in Activation Buffer.
- Dilute Lysyl-Endopeptidase to 0.8 µg/mL in Activation Buffer.
- Activate rhKLK13 by adding 1 µL of 0.8 µg/mL Lysyl-Endopeptidase to 40 µL of 100 µg/mL rhKLK13 for a final concentration of 0.02 µg/mL for Lysyl-Endopeptidase.
- Incubate at 37 °C for 30 minutes.
- Dilute activated rhKLK13 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 µL of 2 ng/µL rhKLK13 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 uM 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).
- rhKLK13: 0.1 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: Kallikrein 13
Human tissue Kallikrein 13 (hK13) is a member of the human tissue kallikrein family. The gene encoding for recombinant hK13 is termed KLK13 and it resides on chromosome 19q13.3-4 along with fourteen other members of the family. KLK13 spans about 8.7 kb genomic DNA and the longest transcript is 831 bp, encoding for a protein of 277 amino acids (1). Another five shorter splice variants have also been identified. They are specifically expressed in the testicular tissue and encode for five truncated forms of hK13 (2). Due to the aspartic acid residue in the substrate binding pocket, the enzymatic activity of hK13 is predicted be trypsin-like. It has been shown that recombinant hK13 produced in yeast can cleave synthetic peptides after the arginine residue and some extracellular matrix components (3). However, its exact physiological substrates and functions remain obscure. Despite the lack of knowledge on the physiological function of hK13, several studies have demonstrated that hK13 is implicated with cancer of the breast and ovary and it can serve as a favorable prognostic biomarker for these malignancies (4, 5). The purified, secreted recombinant hK13 is mainly the pro form. When activated by lysyl‑endopeptidase, it displays enzymatic activity towards a fluorogenic synthetic peptide described in the Activity Assay Protocol. This activity can be inhibited by recombinant human Serpin A5, E1 and F2 (Catalog # 1266-PI, 1786-PI, and 1470‑PI).
- Yousef, G. M. et al. (2000) J. Biol. Chem. 275:11891.
- Chang, A. et al. (2001) Anticancer Res. 21:3147.
- Kapadia, C. et al. (2004) Biochem. Biophys. Res. Commun. 323:1084.
- Chang, A. et al. (2002) Br. J. Cancer. 86:1457.
- Scorilas, A. et al. (2004) J. Clin. Oncol. 22:678.
Citations for Recombinant Human Kallikrein 13 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.
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Citations: Showing 1 - 3
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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: 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 -
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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