Recombinant Human Cathepsin D Protein, CF

R&D Systems | Catalog # 1014-AS

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Cathepsin D Protein (1014-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

P07339.1

Structure / Form

Pro form

Applications

Enzyme Activity
View all Cathepsin D Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Cathepsin D protein
Leu21-Leu412, 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

Leu21

Predicted Molecular Mass

44 kDa

SDS-PAGE

50 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 >350 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 4 using 1014-AS in the following applications:

Formulation, Preparation, and Storage

1014-AS
Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
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 opening.

Background: Cathepsin D

Cathepsin D is a lysosomal aspartic protease of the pepsin family (1). Human cathepsin D is synthesized as a precursor protein, consisting of a signal peptide (residues 1‑18), a propeptide (residues 19‑64), and a mature chain (residues 65‑412) (2‑4). The mature chain can be processed further to the light (residues 65‑161) and heavy (residues 169‑412) chains. 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 are responsible for the generation of angiostatin, a potent endogeneous inhibitor of angiogenesis (6).

References

  1. Conner et al. in Handbook of Proteolytic Enzymes Barrett (1998) Academic Press, San Diego, p. 828.
  2. Faust, et al. (1985) Proc. Natl. Acad. Sci. USA 82:4910.
  3. Westley and May (1987) Nucl. Acid Res. 15:3773.
  4. Redecker, et al. (1991) DNA Cell Biol. 10:423.
  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

P07339.1

Additional Cathepsin D Products

Product Documents for Recombinant Human 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 Human Cathepsin D Protein, CF

For research use only

Citations for Recombinant Human Cathepsin D Protein, CF

Customer Reviews for Recombinant Human Cathepsin D Protein, CF (1)

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Protocols

View specific protocols for Recombinant Human Cathepsin D Protein, CF (1014-AS):

Materials
  • Assay Buffer: 0.1 M NaOAc, 0.2 M NaCl, pH 3.5
  • Recombinant Human Cathepsin D (rhCathepsin D) (Catalog # 1014-AS)
  • Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCathepsin D to 20 µg/mL in Assay Buffer.
  2. Aliquot 50 µL of 20 µg/mL rhCathepsin D.
  3. Incubate at 37 °C for 30 minutes.
  4. Dilute incubated rhCathepsin D to 1 ng/µL in Assay Buffer.
  5. Dilute Substrate to 60 µM in Assay Buffer.
  6. Load 50 µL of 1 ng/µL rhCathepsin D in a plate, and start the reaction by adding 50 µL of 60 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 60 µM Substrate.
  7. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  8. 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:

  • rhCathepsin D: 0.050 µg
  • Substrate: 30 µM

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