Recombinant Human Cathepsin D Protein, CF Summary
Product Specifications
Leu21-Leu412, 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.
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.
|
Assay Procedure
- 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
- Dilute rhCathepsin D to 20 µg/mL in Assay Buffer.
- Aliquot 50 µL of 20 µg/mL rhCathepsin D.
- Incubate at 37 °C for 30 minutes.
- Dilute incubated rhCathepsin D to 1 ng/µL in Assay Buffer.
- Dilute Substrate to 60 µM in Assay Buffer.
- 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.
- 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).
Per Well:- rhCathepsin D: 0.050 µg
- Substrate: 30 µM
Reconstitution Calculator
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).
- Conner et al. in Handbook of Proteolytic Enzymes Barrett (1998) Academic Press, San Diego, p. 828.
- Faust, et al. (1985) Proc. Natl. Acad. Sci. USA 82:4910.
- Westley and May (1987) Nucl. Acid Res. 15:3773.
- Redecker, et al. (1991) DNA Cell Biol. 10:423.
- Rochefort, et al. (2000) Clin. Chim. Acta. 291:157.
- Tsukuba, et al. (2000) Mol. Cells 10:601.
Citations for Recombinant Human 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 - 10
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A New Cathepsin D Targeting Drug Delivery System Based on Immunoliposomes Functionalized with Lipidated Pepstatin A
Authors: Kozak, A;Mikhaylov, G;Khodakivskyi, P;Goun, E;Turk, B;Vasiljeva, O;
Pharmaceutics
Species: N/A
Sample Types: Peptide
Applications: Bioassay -
Intracellular complement C5a/C5aR1 stabilizes beta-catenin to promote colorectal tumorigenesis
Authors: P Ding, Y Xu, L Li, X Lv, L Li, J Chen, D Zhou, X Wang, Q Wang, W Zhang, T Liao, QH Ji, QY Lei, W Hu
Cell Reports, 2022-05-31;39(9):110851.
Species: Human
Sample Types: Recombinant Proteins
Applications: Bioassay -
A topological refactoring design strategy yields highly stable granulopoietic proteins
Authors: J Skokowa, B Hernandez, M Coles, M Ritter, M Nasri, J Haaf, N Aghaallaei, Y Xu, P Mir, AC Krahl, KW Rogers, K Maksymenko, B Bajoghli, K Welte, AN Lupas, P Müller, M ElGamacy
Nature Communications, 2022-05-26;13(1):2948.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
The lysosomal protein cathepsin L is a progranulin protease
Authors: CW Lee, JN Stankowski, J Chew, CN Cook, YW Lam, S Almeida, Y Carlomagno, KF Lau, M Prudencio, FB Gao, M Bogyo, DW Dickson, L Petrucelli
Mol Neurodegener, 2017-07-25;12(1):55.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Oxidation of protein-bound methionine in Photofrin-photodynamic therapy-treated human tumor cells explored by methionine-containing peptide enrichment and quantitative proteomics approach
Authors: YJ Hsieh, KY Chien, IF Yang, IN Lee, CC Wu, TY Huang, JS Yu
Sci Rep, 2017-05-02;7(1):1370.
Applications: Bioassay -
Global protease activity profiling provides differential diagnosis of pancreatic cysts
Authors: SL Ivry, JM Sharib, DA Dominguez, N Roy, SE Hatcher, M Yip-Schnei, CM Schmidt, RE Brand, WG Park, M Hebrok, G Kim, AJ O'Donoghue, KS Kirkwood, CS Craik
Clin. Cancer Res., 2017-04-19;0(0):.
Applications: Bioassay -
Cleavage of Histone 3 by Cathepsin D in the involuting mammary gland.
Authors: Khalkhali-Ellis, Zhila, Goossens, William, Margaryan, Naira V, Hendrix, Mary J C
PLoS ONE, 2014-07-23;9(7):e103230.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
Cathepsin D is partly endocytosed by the LRP1 receptor and inhibits LRP1-regulated intramembrane proteolysis.
Authors: Derocq D, Prebois C, Beaujouin M, Laurent-Matha V, Pattingre S, Smith GK, Liaudet-Coopman E
Oncogene, 2011-11-14;31(26):3202-12.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Regulation of cathepsin g reduces the activation of proinsulin-reactive T cells from type 1 diabetes patients.
Authors: Zou F, Schafer N, Palesch D, Brucken R, Beck A, Sienczyk M, Kalbacher H, Sun Z, Boehm BO, Burster T
PLoS ONE, 2011-08-05;6(8):e22815.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
BACE1 and BACE2 enzymatic activities in Alzheimer's disease.
Authors: Ahmed RR, Holler CJ, Webb RL
J. Neurochem., 2009-12-04;112(4):1045-53.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
Cystatin M/E is a high affinity inhibitor of cathepsin V and cathepsin L by a reactive site that is distinct from the legumain-binding site. A novel clue for the role of cystatin M/E in epidermal cornification.
Authors: Cheng T, Hitomi K, van Vlijmen-Willems IM, de Jongh GJ, Yamamoto K, Nishi K, Watts C, Reinheckel T, Schalkwijk J, Zeeuwen PL
J. Biol. Chem., 2006-03-24;281(23):15893-9.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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