Recombinant Mouse Cathepsin C/DPPI Protein, CF

Catalog # Availability Size / Price Qty
1034-CY-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse 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 >8,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse Cathepsin C/DPPI protein
Asp25-Leu462, with a C-terminal 10-His tag
Accession # P97821
Accession #
N-terminal Sequence
Analysis
Asp25
Structure / Form
Pro form
Predicted Molecular Mass
51 kDa
SDS-PAGE
52 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.

1034-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.
  • 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
  • Activation Buffer: 25 mM MES, 5 mM DTT, pH 5.5
  • Assay Buffer: 25 mM MES, 50 mM NaCl, 5 mM DTT, pH 6.0
  • Recombinant Mouse Cathepsin C/DPPI (rmCathepsin C) (Catalog # 1034-CY)
  • Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
  • Substrate: Gly-Arg-AMC (Bachem, Catalog # I-1215)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmCathepsin C to 20 µg/mL in Activation Buffer.
  2. Dilute rhCathepsin L to 2 µg/mL in Activation Buffer.
  3. Combine equal volumes of diluted rmCathepsin C and diluted rhCathepsin L for final concentrations of 10 µg/mL rmCathepsin C and 1 µg/mL for rhCathepsin L.
  4. Incubate at room temperature for 20 minutes to activate rmCathepsin C.
  5. Dilute activated rmCathepsin C to 0.04 ng/µL in Assay Buffer.
  6. Dilute Substrate to 20 µM in Assay Buffer.
  7. Load 50 µL of the 0.04 ng/µL rmCathepsin C in a 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.
  8. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. 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 (Sigma, Catalog # A9891).

Per Well:
  • rmCathepsin C: 0.002 µg
  • Substrate: 10 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 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).

References
  1. Turk, B. et al. (2004) in Handbook of Proteolytic Enzymes. Barrett, et al. eds. p. 1192, Academic Press, San Diego.
  2. Dahl, S.W. 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

Citation for Recombinant Mouse 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.

1 Citation: Showing 1 - 1

  1. Coxiella burnetii-infected NK cells release infectious bacteria by degranulation
    Authors: S Matthiesen, L Zaeck, K Franzke, R Jahnke, C Fricke, M Mauermeir, S Finke, A Lührmann, MR Knittler
    Infect. Immun., 2020;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Cell Culture

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