Recombinant Mouse Cathepsin D Protein, CF

R&D Systems | Catalog # 1029-AS

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Cathepsin D Protein (1029-AS)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

Q3UCD9

Structure / Form

Pro form

Applications

Enzyme Activity
View all Cathepsin D Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Cathepsin D protein
Ile21-Leu410, with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

Ile21

Predicted Molecular Mass

44 kDa

SDS-PAGE

52 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).
The specific activity is >600 pmol/min/µg, as measured under described conditions.

Formulation, Preparation, and Storage

1029-AS
Formulation Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Reconstitution

Reconstitute at 100 μg/mL in sterile, deionized water.
Shipping The product is shipped with polar packs. 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 reconstitution.

Calculators

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 D

Cathepsin D is a lysosomal aspartic protease of the pepsin family (4). Mouse Cathepsin D is synthesized as a precursor protein, consisting of a signal peptide (residues 1‑20), a propeptide (residues 21‑64), and a mature chain (residues 65‑410) (1‑3). It is expressed in most cells and overexpressed in breast cancer cells (5). It is a major enzyme in protein degradation in lysosomes, and also involved in the presentation of antigenic peptides. Mice deficient in this enzyme showed a progressive atrophy of the intestinal mucosa, a massive destruction of lymphoid organs, and a profound neuronal ceroid lipofucinosis, indicating that Cathepsin D is essential for proteolysis of proteins regulating cell growth and tissue homeostasis (6). Cathepsin D secreted from human prostate carcinoma cells is responsible for the generation of angiostatin, a potent endogeneous inhibitor of angiogenesis (6).

References

  1. Diedrich, et al. (1990) Nucl. Acid Res. 18:7184.
  2. Grusby, et al. (1990) Nucl. Acid Res. 18:4008. 
  3. Hetman, et al. (1994) DNA Cell Biol. 13:419.
  4. Conner (2004) in Handbook of Proteolytic Enzymes (Barrett, et al. eds) Elsevier Academic Press, San Diego, p. 43.
  5. Rochefort, et al. (2000) Clin. Chim. Acta. 291:157.
  6. Tsukuba, et al. (2000) Mol. Cells 10:601.

Alternate Names

CTSD

Entrez Gene IDs

1509 (Human); 13033 (Mouse)

Gene Symbol

CTSD

UniProt

Q3UCD9

Additional Cathepsin D Products

Product Documents for Recombinant Mouse Cathepsin D Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Cathepsin D Protein, CF

For research use only

Citations for Recombinant Mouse Cathepsin D Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Cathepsin D Protein, CF (1029-AS):

Materials
  • Assay Buffer: 0.1 M Sodium Acetate, 0.2 M NaCl, pH 3.5
  • Recombinant Mouse Cathepsin D (rmCathepsin D) (Catalog # 1029-AS)
  • Substrate: MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Catalog # ES001)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmCathepsin D to 20 µg/mL in Assay Buffer.
  2. Incubate at RT for 10 minutes.
  3. Dilute activated rmCathepsin D to 0.4 ng/µL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load 50 µL of the 0.4 ng/µL rmCathepsin D 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.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  7. 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975)

Per Well:

  • rmCathepsin D: 0.020 µg
  • Substrate: 10 µM

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