Cathepsin D Antibody [Unconjugated]

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1014) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). Specific bands were detected for Procathepsin D at approximately 45 kDa and Cathepsin D heavy chain 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western blot shows lysates of PANC‑1 human pancreatic carcinoma cell line and MCF‑7 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1014) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017).Specific bands were detected for Procathepsin D at approximately 45 kDa and Cathepsin D heavy chain 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line and MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. Specific bands were detected for Procathepsin D at approximately 55 kDa and Cathepsin D heavy chain at approximately 37 kDa (as indicated) using 50 µg/mL of Goat Anti-Human Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1014) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Cathepsin D was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Goat Anti-Human Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1014) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, IP
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Cathepsin D Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Cathepsin D
  Leu21-Leu412
  Accession # P07339
• Specificity: Detects human Cathepsin D in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant mouse (rm) Cathepsin D is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CTSD

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Simple Western 50 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Immunoprecipitation 25 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for Cathepsin D Antibody [Unconjugated]

• 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 (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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Cathepsin D Antibody (AF1014)(6)

Publications using AF1014

• X Gao, A De Mazière, DB Iaea, CP Arthur, J Klumperman, C Ciferri, RN Hannoush Visualizing the cellular route of entry of a cystine-knot peptide with Xfect transfection reagent by electron microscopy Sci Rep, 2019;9(1):6907. 2019 [PMID: 31061420] (Electron Microscopy, Human)
• J Han, LA Goldstein, W Hou, S Chatterjee, TF Burns, H Rabinowich HSP90 inhibition targets autophagy and induces a CASP9-dependent resistance mechanism in NSCLC Autophagy, 2018;0(0):1-14. 2018 [PMID: 29561705] (Western Blot, Human)
• Amphiphilic star PEG-Camptothecin conjugates for intracellular targeting J Control Release, 2016;0(0):. 2016 [PMID: 27677603] (IHC-Fr, Human)
• The Apaf-1-binding protein Aven is cleaved by Cathepsin D to unleash its anti-apoptotic potential. Cell Death Differ., 2012;19(9):1435-45. 2012 [PMID: 22388353] (Western Blot, Human)
• Khalkhali-Ellis Z, Abbott DE, Bailey CM, Goossens W, Margaryan NV, Gluck SL, Reuveni M, Hendrix MJ IFN-gamma regulation of vacuolar pH, cathepsin D processing and autophagy in mammary epithelial cells. J. Cell. Biochem., 2008;105(1):208-18. 2008 [PMID: 18494001] (Western Blot, Human)
• Deborde S, Perret E, Gravotta D, Deora A, Salvarezza S, Schreiner R, Rodriguez-Boulan E Clathrin is a key regulator of basolateral polarity. Nature, 2008;452(7188):719-23. 2008 [PMID: 18401403] (Immunoprecipitation, Canine)

Bioinformatics

• Gene Symbol: CTSD
• Uniprot:
  • P07339()