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 # http://www.rndsystems.com/product_results.aspx?k=1266-PI">1266-PI, http://www.rndsystems.com/product_results.aspx?k=1786-PI">1786-PI, and http://www.rndsystems.com/product_results.aspx?k=1470-PI">1470‑PI).
Recombinant Human Kallikrein 13 Protein, CF
R&D Systems | Catalog # 2625-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Kallikrein 13 Protein (2625-SE)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Structure / Form
Pro and mature forms
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 13 protein
Met1-Ile262, with a C-terminal 6-His tag
Met1-Ile262, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Val19 & Val26
Predicted Molecular Mass
28 kDa (pro) & 27 kDa (mature)
SDS-PAGE
31-37 kDa, reducing conditions
Activity
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.
Reviewed Applications
Read 2 reviews rated 5 using 2625-SE in the following applications:
Formulation, Preparation, and Storage
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.
|
Background: Kallikrein 13
References
- 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.
Alternate Names
KLK13
Entrez Gene IDs
26085 (Human)
Gene Symbol
KLK13
UniProt
Additional Kallikrein 13 Products
Product Documents for Recombinant Human Kallikrein 13 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Kallikrein 13 Protein, CF
For research use only
Citations for Recombinant Human Kallikrein 13 Protein, CF
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5 out of 5
2 Customer Ratings
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Application: Immunoassay StandardVerified Customer | Posted 07/05/2022good standard for ELISA
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Protocols
View specific protocols for Recombinant Human Kallikrein 13 Protein, CF (2625-SE):
Materials
- 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
- Substrate: Boc-Val-Pro-Arg-AMC (Catalog # ES011), 100 mM stock in DMSO
- Black 96-well Plate
- Plate Reader with Fluorescence Read Capability
- 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).
Per Well:
- rhKLK13: 0.1 µg
- Substrate: 100 µM