Recombinant Human Chymotrypsin C/CTRC Protein, CF
Recombinant Human Chymotrypsin C/CTRC Protein, CF Summary
Product Specifications
Met1-Leu268, 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.
6907-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
- Assay Buffer: 25 mM Tris, 0.5 mM CaCl2, pH 8.0
- Recombinant Human Chymotrypsin C/CTRC (rhCTRC) (Catalog # 6907-SE)
- Trypsin (Sigma, Catalog # T1426)
- Recombinant Human Serpin F2/ alpha 2‑Antiplasmin (rhSerpin F2) (Catalog # 1470-PI) (optional)
- Substrate: Suc-AAPF-AMC (Bachem, Catalog # I-1465), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhCTRC to 200 µg/mL in Activation Buffer.
- Dilute Trypsin to 0.4 µg/mL in Activation Buffer.
- Combine equal volumes of 200 µg/mL rhCTRC and 0.4 µg/mL Trypsin.
- Incubate at 37 °C for 1 hour.
- Optional: Stop Trypsin activity by adding rhSerpin F2 to a final concentration of 4 µg/mL.
- Dilute rhCTRC to 1 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load into a black well plate 50 µL of the 1 ng/µL rhCTRC and start the reaction by adding 50 µL of 200 µM Substrate. For a Substrate Blank, load Assay Buffer in place of the rhCTRC.
- 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 # A9891).
- rhCTRC: 0.050 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: Chymotrypsin C/CTRC
Chymotrypsin C (CTRC), also known as caldecrin, is a serine protease and a member of the peptidase S1 family. The CTRC gene encodes a serum calcium‑decreasing factor that has chymotrypsin-like protease activity (1). CTRC has broad substrate specificity, but prefers to cleave on the carboxyl side of hydrophobic residues. The enzyme is expressed primarily in the pancreas, and is secreted into the digestive tract. CTRC is a key regulator of activation and degradation of cationic trypsinogen (2, 3). The inappropriate activation of cationic trypsinogen can contribute to the development of pancreatitis (2, 3). CTRC inhibits osteoclast differentiation by suppression of NFATc1 function by a mechanism independent of its proteolytic activity (4).
- Tomomura, A. et al. (1995) J. Biol. Chem. 270:30315.
- Szmola, R. and M. Sahin-Toth (2007) Proc. Natl. Acad. Sci. USA 104:11227.
- Nemoda, Z. and M. Sahin-Toth (2006) J. Biol. Chem. 281:11879.
- Hasegawa, H. et al. (2010) J. Biol. Chem. 285:25448.
Product Specific Notices
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.Citation for Recombinant Human Chymotrypsin C/CTRC 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.
1 Citation: Showing 1 - 1
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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;22(11):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
FAQs
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