Overview

  • Product name

    Anti-Cathepsin D antibody [EPR3056Y]
    See all Cathepsin D primary antibodies
  • Description

    Rabbit monoclonal [EPR3056Y] to Cathepsin D
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICCmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cathepsin D aa 50-150 (N terminal). The exact sequence is proprietary.

  • Positive control

    • MCF-7 and SKBR3 cell lysates. Human liver tissue. WB: A431, HepG2 and MCF7 whole cell lysate
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

Applications

Our Abpromise guarantee covers the use of ab75811 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.
WB 1/1000 - 1/2000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC 1/100 - 1/250.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
    • Tissue specificity

      Expressed in the aorta extrcellular space (at protein level).
    • Involvement in disease

      Ceroid lipofuscinosis, neuronal, 10
    • Sequence similarities

      Belongs to the peptidase A1 family.
      Contains 1 peptidase A1 domain.
    • Post-translational
      modifications

      N- and O-glycosylated.
    • Cellular localization

      Lysosome. Melanosome. Secreted, extracellular space. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. In aortic samples, detected as an extracellular protein loosely bound to the matrix (PubMed:20551380).
    • Information by UniProt
    • Database links

    • Alternative names

      • CatD antibody
      • CATD_HUMAN antibody
      • Cathepsin D antibody
      • Cathepsin D heavy chain antibody
      • CD antibody
      • Ceroid lipofuscinosis neuronal 10 antibody
      • CLN10 antibody
      • CPSD antibody
      • ctsd antibody
      • Epididymis secretory sperm binding protein Li 130P antibody
      • HEL S 130P antibody
      • Lysosomal aspartyl peptidase antibody
      • Lysosomal aspartyl protease antibody
      • MGC2311 antibody
      see all

    Images

    • All lanes : Anti-Cathepsin D antibody [EPR3056Y] (ab75811) at 1/1000 dilution

      Lane 1 : Wild-type A431 whole cell lysate
      Lane 2 : CTSD knockout A431 whole cell lysate
      Lane 3 : HepG2 whole cell lysate
      Lane 4 : MCF7 whole cell lysate

      Lysates/proteins at 40 µg per lane.

      Predicted band size: 44 kDa
      Observed band size: 44 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab75811 observed at 44 kDa. Red - loading control, ab7291, observed at 50 kDa.

      ab75811 was shown to specifically react with in wild-type A431 cells as signal was lost in CTSD knockout cells. Wild-type and CTSD knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab75811 and ab7291 (Mouse anti Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-Cathepsin D antibody [EPR3056Y] (ab75811) at 1/1000 dilution

      Lane 1 : MCF-7 cell lysate
      Lane 2 : SKBR3
      cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 44 kDa
      Observed band size: 15,46 kDa
      why is the actual band size different from the predicted?

    • Immunohistochemical analysis of paraffin-embedded human liver using ab75811 at 1/500 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • ab75811 staining Cathepsin D in the human cell line MCF7 (Human breast adenocarcinoma epithelial cell) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody. Isoytype control: Rabbit monoclonal IgG (Black) Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    References

    This product has been referenced in:

    • Ashraf Y  et al. Immunotherapy of triple-negative breast cancer with cathepsin D-targeting antibodies. J Immunother Cancer 7:29 (2019). Read more (PubMed: 30717773) »
    See 1 Publication for this product

    Customer reviews and Q&As

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (small intestine, ganglion cells)
    Specification
    small intestine, ganglion cells
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate, 100C 20 minutes
    Permeabilization
    No

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    Verified customer

    Submitted Oct 22 2009

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