Mouse Cathepsin D Antibody

R&D Systems | Catalog # AF1029

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Cathepsin D Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Cathepsin D
Ile21-Leu410
Accession # Q3UCD9

Specificity

Detects mouse Cathepsin D in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse Cathepsin D Antibody

Detection of Mouse Cathepsin D antibody by Western Blot.

Detection of Mouse Cathepsin D by Western Blot.

Western blot shows lysate of mouse intestine tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1029) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). Specific bands were detected for Cathepsin D at approximately 28 and 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Cathepsin D antibody in RAW 264 by Immunocytochemistry (ICC).7 Mouse Cell Line by Immunocytochemistry (ICC).

Cathepsin D in RAW 264.7 Mouse Cell Line.

Cathepsin D was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Goat Anti-Mouse Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1029) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to lysosomes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Cathepsin D antibody in Mouse Intestine by Immunohistochemistry (IHC-Fr).

Cathepsin D in Mouse Intestine.

Cathepsin D was detected in perfusion fixed frozen sections of mouse intestine using 15 µg/mL Goat Anti-Mouse Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1029) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counter-stained with hematoxylin (blue). Specific labeling was localized to epithelial cells in intestinal villi. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of Human Cathepsin D by Western Blot

Detection of Human Cathepsin D by Western Blot

Chronic treatment with BACE1 inhibitors causes accumulation of CatD.(a) Representative western blot of brain homogenates prepared from mice dosed daily for 30 days with PF-9283 (80 mg kg−1) showed increased levels of CatD heavy and light chains, but not the CatD single chain, compared with a vehicle control. Data are presented as mean (normalized to actin)±s.e.m.; n=5 (PF-9283) and n=6 (veh) per group. (b) ELISA of live ARPE-19 cells incubated with BACE1 inhibitors for 7 days showed accumulation of CatD; the extent of accumulation was consistent with cellular CatD IC50 values (Supplementary Table 4). Data are presented as mean±s.e.m. for three independent experiments. (c) Proteomic analysis of RPE tissue prepared from rats dosed daily for 14 days with 80 mg kg−1 PF-9283. Peptide spectral matches (PSM) of 11 of 391 identified proteins were changed more than 1.5-fold (P<0.01, red symbols, see also Supplementary Table 5) compared with vehicle controls including a 1.7-fold increase in CatD (***P=0.0002). Average PSM were calculated from six LC-MS runs consisting of two technical replicates of n=3 RPE tissues per group. For a–c, significant differences were determined using a two-tailed t-test; *P<0.05, **P<0.01, ***P<0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27727204), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Cathepsin D by Western Blot

Detection of Human Cathepsin D by Western Blot

Chronic treatment with BACE1 inhibitors causes accumulation of CatD.(a) Representative western blot of brain homogenates prepared from mice dosed daily for 30 days with PF-9283 (80 mg kg−1) showed increased levels of CatD heavy and light chains, but not the CatD single chain, compared with a vehicle control. Data are presented as mean (normalized to actin)±s.e.m.; n=5 (PF-9283) and n=6 (veh) per group. (b) ELISA of live ARPE-19 cells incubated with BACE1 inhibitors for 7 days showed accumulation of CatD; the extent of accumulation was consistent with cellular CatD IC50 values (Supplementary Table 4). Data are presented as mean±s.e.m. for three independent experiments. (c) Proteomic analysis of RPE tissue prepared from rats dosed daily for 14 days with 80 mg kg−1 PF-9283. Peptide spectral matches (PSM) of 11 of 391 identified proteins were changed more than 1.5-fold (P<0.01, red symbols, see also Supplementary Table 5) compared with vehicle controls including a 1.7-fold increase in CatD (***P=0.0002). Average PSM were calculated from six LC-MS runs consisting of two technical replicates of n=3 RPE tissues per group. For a–c, significant differences were determined using a two-tailed t-test; *P<0.05, **P<0.01, ***P<0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27727204), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Cathepsin D by Immunocytochemistry/ Immunofluorescence

Detection of Cathepsin D by Immunocytochemistry/ Immunofluorescence

sPLA2 inhibitor treatment rescues PGRN deficiency phenotypes in mice. 5-week-old Pla2g2a+/+Grn+/+ and Pla2g2a+/+Grn−/− mice in FVB/B6 mixed background were treated with corn oil control or sPLA2 inhibitors, LY333013 or KH064 for 3 weeks. (a) Measurement of total sPLA2 activity in the serum and cortical lysates from untreated and LY333013 or KH064-treated mice. Data are presented as mean ± SEM from five mice per group (n = 5). *, p < 0.05; **, p < 0.01, Student’s t-test. (b) Brain sections were immunostained with anti-GFAP, CD68, and IBA1 antibodies, and representative images from the cortex region were shown (Scale bar = 100 μm). Another set of brain sections was stained with Hoechst and auto-fluorescent lipofuscin signals (red) in the thalamus region were imaged (scale bar = 10 μm). (c) Quantification of GFAP, CD68, and IBA1 levels and auto-fluorescent lipofuscin signals for the experiment in (b). Data are presented as mean ± SEM from five mice per group (n = 5). 3 sections were analyzed for each mouse brain. *, p < 0.05; **, p < 0.01, Student’s t-test. (d) Brain sections were immunostained with anti-CathD, NeuN, and IBA1 antibodies, and representative images from the cortex region were shown (Scale bar = 10 μm). (e) CathD signals in IBA1-positive microglia and NeuN-positive neurons were quantified. 10–20 confocal images were randomly captured from each brain section. Ten to twenty microglia and fifty to sixty neurons were quantified per section. 3 sections were analyzed for each mouse brain. Data are presented as mean ± SEM from 3–4 mice per group (n = 3–4). *, p < 0.05; **, p < 0.01, Student’s t-test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40528203), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Cathepsin D by Immunocytochemistry/ Immunofluorescence

Detection of Cathepsin D by Immunocytochemistry/ Immunofluorescence

AAV9-mediated overexpression of sPLA2-IIA leads to lipofuscin accumulation and lysosomal enlargement in microglia in 2-month-old B6 Grn−/− mice. B6 WT (Pla2g2−/−Grn+/+) and Grn−/− (Pla2g2a−/−Grn−/−) pups at postnatal day (P0) were injected with AAV9-sPLA2-IIA or the buffer control, and the mice were collected at 2 months old. (a) Brain sections were stained with NeuN and IBA1, and auto-fluorescent lipofuscin signals (blue) in the cortex region were imaged (scale bar = 10 μm). (b) Quantification of autofluorescent lipofuscin signals in the cortex. Data are presented as mean ± SEM from three to five mice per group (n = 3–5). Three sections were analyzed for each mouse brain. *, p < 0.05; **, p < 0.01; ****, p < 0.0001, Student’s t-test. (c) Brain sections were immunostained with anti-CathD, NeuN, and IBA1 antibodies, and representative images from the cortex region were shown (Scale bar = 10 μm). (d) CathD signals in IBA1-positive microglia and NeuN-positive neurons were quantified. 10–20 confocal images were randomly captured from each brain section. Fifteen to thirty microglia and fifty to sixty neurons were quantified per Sect. 3 sections were analyzed for each mouse brain. Data are presented as mean ± SEM from 3–5 mice per group (n = 3–5). *, p < 0.05; **, p < 0.01; ****, p < 0.0001, Student’s t-test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40528203), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse Cathepsin D Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed RAW264.7 mouse monocyte/macrophage cell line

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse intestine

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Mouse Cathepsin D (Catalog # 1029-AS), see our available Western blot detection antibodies

Western Blot

1 µg/mL
Sample:   Mouse intestine tissue

Reviewed Applications

Read 2 reviews rated 5 using AF1029 in the following applications:

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.

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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 over-expressed 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).

References

  1. Diedrich, et al. (1990) Nucl. Acid Res. 18:7184.
  2. Grusby, et al. (1990) Nucl. Acid Res. 18:4008. 
  3. Hetman, et al. (1994) DNA Cell Biol. 13:419.
  4. Conner, (2004) in Handbook of Proteolytic Enzymes (Barrett, et al. eds) Elsevier Academic Press, San Diego, p. 43.
  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

Q3UCD9

Additional Cathepsin D Products

Product Documents for Mouse Cathepsin D Antibody

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Product Specific Notices for Mouse Cathepsin D Antibody

For research use only

Citations for Mouse Cathepsin D Antibody

Customer Reviews for Mouse Cathepsin D Antibody (2)

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  • Mouse Cathepsin D Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Colon tissue
    Species: Mouse
    Verified Customer | Posted 08/05/2020
    1:200 dilution
    Mouse Cathepsin D Antibody AF1029
  • Mouse Cathepsin D Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Bladder tissue
    Species: Human
    Verified Customer | Posted 10/31/2017

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