Recombinant Human Kallikrein 13 Protein, CF

Catalog # Availability Size / Price Qty
2625-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (3)
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Recombinant Human Kallikrein 13 Protein, CF Summary

Product Specifications

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.
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.
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 13 protein
Met1-Ile262, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Val19 & Val26
Structure / Form
Pro and mature forms
Predicted Molecular Mass
28 kDa (pro) & 27 kDa (mature)
SDS-PAGE
31-37 kDa, reducing conditions

Product Datasheets

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.

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.
  • 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: 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
  1. Dilute rhKLK13 to 100 µg/mL in Activation Buffer.
  2. Dilute Lysyl-Endopeptidase to 0.8 µg/mL in Activation Buffer.
  3. 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.
  4. Incubate at 37 °C for 30 minutes.
  5. Dilute activated rhKLK13 to 2 ng/µL in Assay Buffer.
  6. Dilute Substrate to 200 µM in Assay Buffer.
  7. 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.
  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:
  • rhKLK13: 0.1 µg
  • Substrate: 100 µM
Reconstitution Calculator

Reconstitution Calculator

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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).

References
  1. Yousef, G. M. et al. (2000) J. Biol. Chem. 275:11891.
  2. Chang, A. et al. (2001) Anticancer Res. 21:3147.
  3. Kapadia, C. et al. (2004) Biochem. Biophys. Res. Commun. 323:1084.
  4. Chang, A. et al. (2002) Br. J. Cancer. 86:1457.
  5. Scorilas, A. et al. (2004) J. Clin. Oncol. 22:678.
Entrez Gene IDs
26085 (Human)
Alternate Names
DKFZp586J1923; EC 3.4.21; EC 3.4.21.-; Kallikrein 13; Kallikrein-like protein 4; kallikrein-related peptidase 13; KLK13; KLK-L4kallikrein-13; KLKL4kallikrein-like gene 4

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.

3 Citations: Showing 1 - 3
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  1. 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;22(11):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  2. 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.
    Species: Human
    Sample Types: Antibody
    Applications: Surface Plasmon Resonance
  3. 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, 0;32(6):1187-92.
    Applications: Bioassay

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Fluorogenic Peptide Substrates

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Recombinant Human Kallikrein 13 Protein, CF
By Anonymous on 04/02/2020
Application: Enzymatic activity in vitro