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).
Human Cathepsin D Antibody
R&D Systems | Catalog # AF1014
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Mouse, Canine, Gerbil, Xenograft
Applications
Validated:
Immunohistochemistry, Western Blot, Simple Western, Immunoprecipitation
Cited:
Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry, Immunoprecipitation, ELISA Capture, Microarray, Electron Microscopy
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Cathepsin D
Leu21-Leu412
Accession # P07339
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.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human Cathepsin D Antibody
Detection of Human Cathepsin D by Western Blot.
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.Detection of Human Cathepsin D by Western Blot.
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.Cathepsin D in Human Lung Cancer Tissue.
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.Detection of Human Cathepsin D by Simple WesternTM.
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.Detection of Cathepsin D by Western Blot
Lysosomal LAMP2A protein levels decrease in POMC-like neurons treated with PA and SA. (A) POMC-like neurons (N43/5) were incubated with 100 mM of PA, SA, or vehicle (BSA) for 6 h and a mitochondrial and lysosomal fractionation was carried out according to Ormeño et al. 2020. Post nuclear (PN), lysosomal (LYS), and mitochondrial (MIT) fractions were evaluated by Western blot using LAMP1, CATD, LAMP2, VDAC, and MTCO1 antibodies, as appropriate. (B) Representative Western blot comparing the lysosomal LAMP2A protein levels versus the PN fractions after BSA, PA, and SA treatment. (C) Quantification of LAMP2A from Figure 2B using ordinary one-way ANOVA (n = 3). n.s. = non-significant, ** = p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35326371), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human Cathepsin D Antibody
Application
Recommended Usage
Immunohistochemistry
5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast and lung cancer tissue
Sample: Immersion fixed paraffin-embedded sections of human breast and lung cancer tissue
Immunoprecipitation
25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Cathepsin D (Catalog # 1014-AS), see our available Western blot detection antibodies
Sample: Conditioned cell culture medium spiked with Recombinant Human Cathepsin D (Catalog # 1014-AS), see our available Western blot detection antibodies
Simple Western
50 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line and MCF‑7 human breast cancer cell line
Sample: K562 human chronic myelogenous leukemia cell line and MCF‑7 human breast cancer cell line
Western Blot
1 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line, PANC‑1 human pancreatic carcinoma cell line, and MCF‑7 human breast cancer cell line
Sample: K562 human chronic myelogenous leukemia cell line, PANC‑1 human pancreatic carcinoma cell line, and MCF‑7 human breast cancer cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & 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.
Calculators
Background: Cathepsin D
References
- 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.
Alternate Names
CTSD
Gene Symbol
CTSD
UniProt
Additional Cathepsin D Products
Product Documents for Human Cathepsin D Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Cathepsin D Antibody
For research use only
Citations for Human Cathepsin D Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Immunoprecipitation Protocol
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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