Recombinant Mouse Cathepsin D Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Mouse Cathepsin D Protein, CF Summary

Details of Functionality
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.
Source
Mouse myeloma cell line, NS0-derived mouse Cathepsin D protein
Ile21-Leu410, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Ile21
Structure / Form
Pro form
Protein/Peptide Type
Recombinant Enzymes
Gene
Ctsd
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
44 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
52 kDa, reducing conditions
Publications
Read Publications using
1029-AS in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile, deionized water.
Assay Procedure
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Cathepsin D Protein, CF

  • cathepsin D (lysosomal aspartyl protease)
  • Cathepsin D
  • CPSD
  • CTSD
  • EC 3.4.23
  • EC 3.4.23.5
  • lysosomal aspartyl peptidase
  • lysosomal aspartyl protease
  • MGC2311
  • neuronal 10

Background

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).

  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.

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Publications for Cathepsin D (1029-AS)(2)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 2 applications: Bioassay, Enzyme Assay.


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Bioassay
(1)
Enzyme Assay
(1)
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Bioinformatics

Gene Symbol Ctsd
Uniprot