Recombinant Human Chymotrypsin C/CTRC Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Chymotrypsin C/CTRC Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, SUC-Ala-Ala-Pro-Phe-AMC. The specific activity is >400 pmol/min/μg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human Chymotrypsin C/CTRC protein
Met1-Leu268, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Cys17
Protein/Peptide Type
Recombinant Enzymes
Gene
CTRC
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
28.8 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
33-40 kDa, reducing conditions
Publications
Read Publication using
6907-SE in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
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
  1. Dilute rhCTRC to 200 µg/mL in Activation Buffer.
  2. Dilute Trypsin to 0.4 µg/mL in Activation Buffer.
  3. Combine equal volumes of 200 µg/mL rhCTRC and 0.4 µg/mL Trypsin.
  4. Incubate at 37 °C for 1 hour.
  5. Optional: Stop Trypsin activity by adding rhSerpin F2 to a final concentration of 4 µg/mL.
  6. Dilute rhCTRC to 1 ng/µL in Assay Buffer.
  7. Dilute Substrate to 200 µM in Assay Buffer.
  8. 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.
  9. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  10. 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:
  • rhCTRC: 0.050 µg
  • Substrate: 100 µM

Notes

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.



This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Chymotrypsin C/CTRC Protein, CF

  • Caldecrin
  • chymotrypsin C (caldecrin)
  • Chymotrypsin C
  • chymotrypsin-C
  • CLCR
  • CLCRserum calcium decreasing factor
  • CTRC
  • EC 3.4.21
  • EC 3.4.21.2
  • ELA4
  • elastase 4
  • Elastase IV

Background

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).
  1. Tomomura, A. et al. (1995) J. Biol. Chem. 270:30315.
  2. Szmola, R. and M. Sahin-Toth (2007) Proc. Natl. Acad. Sci. USA 104:11227.
  3. Nemoda, Z. and M. Sahin-Toth (2006) J. Biol. Chem. 281:11879.
  4. Hasegawa, H. et al. (2010) J. Biol. Chem. 285:25448.

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6907-SE
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Publications for Chymotrypsin C/CTRC (6907-SE)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: Bioassay.


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Bioinformatics

Gene Symbol CTRC
Uniprot