Recombinant Mouse Kallikrein 5 Protein, CF

Catalog # Availability Size / Price Qty
7236-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse Kallikrein 5 Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
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 >1,350 pmol/min/μg, as measured under the described conditions. www.RnDSystems.com.
Source
Mouse myeloma cell line, NS0-derived mouse Kallikrein 5 protein
Val23-Asn293, with a C-terminal 10‑His tag
Accession #
N-terminal Sequence
Analysis
Val23
Predicted Molecular Mass
31 kDa
SDS-PAGE
40-50 kDa, reducing conditions

Product Datasheets

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7236-SE

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.

7236-SE

Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate, 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
  • Assay Buffer: 50 mM Tris, 0.005% (w/v) Brij-35, pH 8.0
  • Recombinant Mouse Kallikrein 5 (rmKLK5) (Catalog # 7236-SE)
  • Recombinant Human Kallikrein 5 (rhKLK5) (Catalog # 1108-SE)
  • Substrate: Boc-Val-Pro-Arg-AMC (Catalog # ES011)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmKLK5 to 200 µg/mL in Assay Buffer.
  2. Dilute rhKLK5 to 2.02 µg/mL in Assay Buffer.
  3. Combine equal volumes of 200 µg/mL rmKLK5 and 2.02 µg/mL rhKLK5.  For a control, combine equal volumes of 2.02 µg/mL rhKLK5 and Assay Buffer.
  4. Incubate at 37 °C for 16 - 20 hours.
  5. Dilute activated rmKLK5 to 1 µg/mL in Assay Buffer.  Perform same dilution on control.
  6. Dilute Substrate to 800 µM in Assay Buffer.
  7. In a plate, load 50 µL of 1 µg/mL rmKLK5 and control, and start the reaction by adding 50 µL of 800 µM Substrate to all wells.
  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 # A9891).

Per Well:
  • rmKLK5: 0.050 µg
  • Substrate: 400 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 5

Tissue Kallikrein 5 (KLK5) is a secreted serine protease with trypsin‑like substrate specificity. KLK5 is highly expressed in the skin, and is also known as stratum corneum tryptic enzyme due to its high expression level in this tissue. KLK5 is thought to play an important role in the process of desquamation (1). KLK5 is a potential biomarker for ovarian and breast cancers (2). It may also be a physiological activator of KLK7 in the stratum corneum (3). Recombinant mouse KLK5 is purified as the proform, and can undergo autoactivation under the appropriate conditions. KLK5 enzymatic activity can be inhibited by the following recombinant human serpins: Serpin‑A1, ‑A5, ‑F2, and ‑G1 (Catalog # 1268-PI, 1266-PI, 1470-PI, and 2488-PI.

References
  1. Brattsand M. and T. Egelrud (1999) J. Biol. Chem. 274:30033.
  2. Yousef G.M. et al. (2003) Cancer Res. 63:3958.
  3. Kim H. et al. (2001) Br. J. Cancer 84:543.
Entrez Gene IDs
25818 (Human); 68668 (Mouse); 690603 (Rat)
Alternate Names
EC 3.4.21; EC 3.4.21.4; Kallikrein 5; Kallikrein-like protein 2; kallikrein-related peptidase 5; KLK5; KLKL2; KLK-L2; KLK-L2EC 3.4.21.-; SCTE; SCTEkallikrein-5; Stratum corneum tryptic enzyme

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