Recombinant Mouse Cathepsin D Protein, CF Summary
Product Specifications
Ile21-Leu410, with a C-terminal 10-His tag
Analysis
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.
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.
|
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
- Dilute rmCathepsin D to 20 µg/mL in Assay Buffer.
- Incubate at RT for 10 minutes.
- Dilute activated rmCathepsin D to 0.4 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 320 nm and 405 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975)
- rmCathepsin D: 0.020 µg
- Substrate: 10 µM
Reconstitution Calculator
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).
- Diedrich, et al. (1990) Nucl. Acid Res. 18:7184.
- Grusby, et al. (1990) Nucl. Acid Res. 18:4008.
- Hetman, et al. (1994) DNA Cell Biol. 13:419.
- Conner (2004) in Handbook of Proteolytic Enzymes (Barrett, et al. eds) Elsevier Academic Press, San Diego, p. 43.
- Rochefort, et al. (2000) Clin. Chim. Acta. 291:157.
- Tsukuba, et al. (2000) Mol. Cells 10:601.
Citations for Recombinant Mouse Cathepsin D 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 - 3
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Cathepsin D interacts with adenosine A2A receptors in mouse macrophages to modulate cell surface localization and inflammatory signaling
Authors: A Skopál, T Kéki, PÁ Tóth, B Csóka, B Koscsó, ZH Német, L Antonioli, A Ivessa, F Ciruela, L Virág, G Haskó, E Kókai
The Journal of Biological Chemistry, 2022-03-31;0(0):101888.
Species: Mouse
Sample Types: Cell Lysates
Applications: Bioassay -
Lysosomal processing of progranulin
Authors: X Zhou, DH Paushter, T Feng, L Sun, T Reinheckel, F Hu
Mol Neurodegener, 2017-08-23;12(1):62.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay -
The cystatin M/E-cathepsin L balance is essential for tissue homeostasis in epidermis, hair follicles, and cornea.
Authors: Zeeuwen PL, van Vlijmen-Willems IM, Cheng T
FASEB J., 2010-05-21;24(10):3744-55.
Species: Mouse
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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