Recombinant Mouse Active Cathepsin C/DPPI Protein, CF

Catalog # Availability Size / Price Qty
2336-CY-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (2)
FAQs
Reviews (2)

Recombinant Mouse Active Cathepsin C/DPPI Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
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, Gly-Arg-7-amido-4-methylcoumarin (GR-AMC). The specific activity is > 60,000 pmol/min/µg, as measured under the described conditions. 
Source
Mouse myeloma cell line, NS0-derived mouse Cathepsin C/DPPI protein
proform (Asp25-Leu462 with a C-terminal 10-His tag)
The proform was activated by Recombinant Human Cathepsin L (Catalog # 952-CY) and further purified.
Accession #
N-terminal Sequence
Analysis
Asp25 (Exclusion domain) and Asp394 (Light chain)
Structure / Form
Active form
Predicted Molecular Mass
13 kDa (Exclusion domain), 18 kDa (Heavy chain), and 9 kDa (Light chain)
SDS-PAGE
8 kDa and 20-27 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.

2336-CY

Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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 MES, 50 mM NaCl, 5 mM DTT, pH 5.5
  • Recombinant Mouse Active Cathepsin C/DPPI (rmCathepsin C) (Catalog # 2336-CY)
  • Fluorogenic Peptide Substrate: Gly-Arg-AMC (Bachem, Catalog # I-1215), 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 rmCathepsin C to 0.01 ng/µL in Assay Buffer.
  2. Dilute Substrate to 800 µM in Assay Buffer.
  3. Load into a black well plate 50 µL of 0.01 ng/µL rmCathepsin C, and start the reaction by adding 50 µL of 800 µM Substrate. Include a Substrate Blank containing Assay Buffer in place of rmCathepsin C.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. 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:
  • rmCathepsin C: 0.0005 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: Cathepsin C/DPPI

Cathepsin C (CTSC) is a cysteine protease of the papain family (1). It sequentially removes dipeptides from the free N-termini of proteins and peptides. It has broad specificity except that it does not cleave a basic amino acid (Arg or Lys) in the N-terminal position or Pro on either side of the scissle bond. It requires halide ions for activity. The pro form contains a pro region and a mature region, which are further cleaved during activation to remove the prodomain and split the catalytic domain into heavy and light chains. The N-terminal domain of the pro region is also called the exclusion domain, which remains connected to the heavy chain through non-covalent bonds to the mature, active enzyme. Broadly distributed, CTSC plays a major role in lysosomal degradation and enzyme activation. For example, it activates granule serine proteases in cytotoxic T lymphocytes and natural killer cells (granzymes A and B), mast cells (tryptase and chymase), and neutrophils (Cathepsin G and elastase) by removing their N-terminal activation dipeptides (2).

References
  1. Turk, et al. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 1192, Academic Press, San Diego.
  2. Dahl, et al. (2001) Biochemistry 40:1671.
Entrez Gene IDs
1075 (Human); 13032 (Mouse); 25423 (Rat)
Alternate Names
Cathepsin C; cathepsin CEC 3.4.14.1; Cathepsin J; CPPIHMS; CTSC; dipeptidyl peptidase 1; Dipeptidyl peptidase I; Dipeptidyl transferase; dipeptidyl-peptidase I; DPP1; DPPI; DPP-I; JP; JPD; PALS; PLS

Citations for Recombinant Mouse Active Cathepsin C/DPPI 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.

2 Citations: Showing 1 - 2
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  1. N-Acetyldopamine dimers from Oxya chinensis sinuosa attenuates lipopolysaccharides induced inflammation and inhibits cathepsin C activity
    Authors: A Bahuguna, TP Khaket, VK Bajpai, S Shukla, I Park, M Na, YS Huh, YK Han, SC Kang, M Kim
    Computational and structural biotechnology journal, 2022;20(0):1177-1188.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Distinct roles for cysteine cathepsin genes in multistage tumorigenesis.
    Authors: Gocheva V, Zeng W, Ke D, Klimstra D, Reinheckel T, Peters C, Hanahan D, Joyce JA
    Genes Dev., 2006;20(5):543-56.
    Species: Mouse
    Sample Types: Buffer
    Applications: Enzyme Assay

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Reviews for Recombinant Mouse Active Cathepsin C/DPPI Protein, CF

Average Rating: 4.5 (Based on 2 Reviews)

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Recombinant Mouse Active Cathepsin C/DPPI Protein, CF
By Anonymous on 12/01/2017
Application: In vivo study

Recombinant Mouse Active Cathepsin C/DPPI Protein, CF
By Anonymous on 09/27/2017
Application: Cell Proliferation