Recombinant Mouse Active Cathepsin C/DPPI Protein, CF
Recombinant Mouse Active Cathepsin C/DPPI Protein, CF Summary
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.
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.
|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.
- 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
- Dilute rmCathepsin C to 0.01 ng/µL in Assay Buffer.
- Dilute Substrate to 800 µM in Assay Buffer.
- 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.
- 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 # A-9891).Per Well:
- rmCathepsin C: 0.0005 µg
- Substrate: 400 µM
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).
- Turk, et al. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 1192, Academic Press, San Diego.
- Dahl, et al. (2001) Biochemistry 40:1671.
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.
Citations: Showing 1 - 2
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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, 2022-02-15;20(0):1177-1188.
Sample Types: Whole Cells
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., 2006-02-15;20(5):543-56.
Sample Types: Buffer
Applications: Enzyme Assay
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