Recombinant Human Cathepsin C/DPPI Protein, CF Summary
Product Specifications
Asp25-Leu463, with a C-terminal 10-His tag
Analysis
51 kDa
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.
1071-CY
| Formulation | Supplied as a 0.2 μm filtered solution in MES and NaCl. |
| 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 Procedure
- Activation Buffer: 25 mM MES, 5 mM DTT, pH 6.0
- Assay Buffer: 25 mM MES, 50 mM NaCl, 5 mM DTT, pH 6.0
- Recombinant Human Cathepsin C/DPPI (rhCathepsin C) (Catalog # 1071-CY)
- Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
- 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 rhCathepsin C to 200 µg/mL in Activation Buffer.
- Add rhCathepsin L and Activation Buffer to rhCathepsin C for final concentrations of 20 µg/mL rhCathepsin L and 100 µg/mL rhCathepsin C.
- Incubate at room temperature for 1 hour.
- Dilute activated rhCathepsin C to 0.5 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 0.5 ng/µL rhCathepsin C into a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rhCathepsin 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).
- rhCathepsin C: 0.025 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: Cathepsin C/DPPI
Cathepsin C, also known as dipeptidyl-peptidase I (DPPI), is a cysteine protease of the papain family (1). Cathepsin C 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 peptide and a catalytic region, which can be further processed into heavy/ alpha and light/ beta chains that are linked by a disulfide bond. It is broadly distributed. Cathepsin C plays a role in the lysosomal degradation. It also functions as a key enzyme in the activation of 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). Loss of function mutations in the Cathepsin C gene result in periodontal disease and palmoplantar keratosis (3).
- Turk, B. et al. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) p. 1192, Academic Press, San Diego.
- Dahl, S.W. et al. (2001) Biochemistry 40:1671.
- Toomes, A.J. et al. (1999) Nat. Genet. 23:421.
Citations for Recombinant Human 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.
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Citations: Showing 1 - 8
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Hederacolchiside A1 Suppresses Autophagy by Inhibiting Cathepsin C and Reduces the Growth of Colon Cancer
Authors: S Kim, KH Lee, HJ Choi, E Kim, S Kang, M Han, HJ Jeon, MY Yun, GY Song, HJ Lee
Cancers, 2023-02-16;15(4):.
Species: N/A
Sample Types: Fluorogenic Peptide Substrate
Applications: Bioassay -
Repression of CTSG, ELANE and PRTN3-mediated histone H3 proteolytic cleavage promotes monocyte-to-macrophage differentiation
Authors: P Cheung, S Schaffert, SE Chang, M Dvorak, M Donato, C Macaubas, MH Foecke, TM Li, L Zhang, JP Coan, GS Schulert, AA Grom, LA Henderson, PA Nigrovic, JE Elias, O Gozani, ED Mellins, P Khatri, PJ Utz, AJ Kuo
Nature Immunology, 2021-05-20;22(6):711-722.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Decursin inhibits cell growth and autophagic flux in gastric cancer via suppression of cathepsin C
Authors: S Kim, SI Lee, N Kim, M Joo, KH Lee, MW Lee, HJ Jeon, H Ryu, JM Kim, JY Sul, GY Song, JY Kim, HJ Lee
American journal of cancer research, 2021-04-15;11(4):1304-1320.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Suppression and resolution of autoimmune arthritis by rhesus ?-defensin-1, an immunomodulatory macrocyclic peptide
Authors: JB Schaal, DQ Tran, A Subramania, R Patel, T Laragione, KD Roberts, K Trinh, P Tongaonkar, PA Tran, D Minond, GB Fields, P Beringer, AJ Ouellette, PS Gulko, ME Selsted
PLoS ONE, 2017-11-16;12(11):e0187868.
Applications: Bioassay -
Cystatin F is a cathepsin C-directed protease inhibitor regulated by proteolysis.
Authors: Hamilton G, Colbert JD, Schuettelkopf AW, Watts C
EMBO J., 2008-02-06;27(3):499-508.
Applications: Enzyme Assay -
Design of protease-resistant myelin basic protein-derived peptides by cleavage site directed amino acid substitutions.
Authors: Burster T, Marin-Esteban V, Boehm BO, Dunn S, Rotzschke O, Falk K, Weber E, Verhelst SH, Kalbacher H, Driessen C
Biochem. Pharmacol., 2007-08-02;74(10):1514-23.
Species: Human
Sample Types: Cell Lysates
Applications: Enzyme Assay -
Inhibition of the activation of multiple serine proteases with a cathepsin C inhibitor requires sustained exposure to prevent pro-enzyme processing.
Authors: Methot N, Rubin J, Guay D, Beaulieu C, Ethier D, Reddy TJ, Riendeau D, Percival MD
J. Biol. Chem., 2007-05-29;282(29):20836-46.
Applications: Enzyme Assay -
An activity-based probe for the determination of cysteine cathepsin protease activities in whole cells.
Authors: Falgueyret JP, Black WC, Cromlish W, Desmarais S, Lamontagne S, Mellon C, Riendeau D, Rodan S, Tawa P, Wesolowski G, Bass KE, Venkatraman S, Percival MD
Anal. Biochem., 2004-12-15;335(2):218-27.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
FAQs
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Average Rating: 3 (Based on 1 Review)
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Reason for Rating: While Cathepsin did activate ms Neutrophil elastase Cat No. 4517-SE-010 the amount of total protein activated was less. The Activity assay of NE after cathepsin activation was not at 100% compared to a control hu NE sample.
I ran the activity assay at greater than 1ng/mL.
