RabMAb

Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

Abreviews (4)Q&A (2)Specific References (4)
Customer reviews and FAQs

Overview

  • Product nameAnti-Cathepsin D antibody [EPR3057Y]
    See all Cathepsin D primary antibodies
  • Description
    Rabbit monoclonal [EPR3057Y] to Cathepsin D
  • Tested applicationsSuitable for: Flow Cyt, ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cathepsin D aa 350 to the C-terminus (C terminal).

  • Positive control
    • WB: MCF-7, A431, SK-BR-3 and HepG2 cell lysates and mouse brain tissue lysate. IHC-P: Human breast carcinoma and liver tissues. ICC/IF: MCF-7 cells. IP: SK-BR-3 cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

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

     

    Alternative versions available:

    Anti-Cathepsin D antibody (Alexa Fluor® 647) [EPR3057Y] (ab198326)

    Anti-Cathepsin D antibody (Alexa Fluor® 488) [EPR3057Y] (ab197605)

     

Properties

Applications

Our Abpromise guarantee covers the use of ab75852 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.
ICC/IF 1/100 - 1/1000.
WB 1/2000 - 1/10000. Predicted molecular weight: 46 kDa.
IP 1/10 - 1/20.
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Target

  • FunctionAcid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
  • Tissue specificityExpressed in the aorta extrcellular space (at protein level).
  • Involvement in diseaseCeroid lipofuscinosis, neuronal, 10
  • Sequence similaritiesBelongs to the peptidase A1 family.
    Contains 1 peptidase A1 domain.
  • Post-translational
    modifications    N- and O-glycosylated.
  • Cellular localizationLysosome. 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

    Anti-Cathepsin D antibody [EPR3057Y] images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Cathepsin D with purified ab75852 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Cathepsin D with purified ab75852 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Anti-Cathepsin D antibody [EPR3057Y] (ab75852) at 1/5000 dilution (purified) + Mouse brain tissue lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Predicted band size : 46 kDa
      Observed band size : 28,43 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM /TBST.

    • Flow Cytometry - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Flow Cytometry - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling Cathepsin D with purified ab75852 at 1/20 dilution (10µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    • Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      All lanes : Anti-Cathepsin D antibody [EPR3057Y] (ab75852) at 1/5000 dilution (purified)

      Lane 1 : MCF-7 whole cell lysate
      Lane 2 : SKBR-3 whole cell lysate
      Lane 3 : HepG2 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Predicted band size : 46 kDa
      Observed band size : 28,43,46 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM /TBST.

      ab75852 detects an extra band at 75kDa.

    • Immunoprecipitation - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Immunoprecipitation - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      ab75852 (purified) at 1/20 immunoprecipitating Cathepsin D in SK-BR-3 whole cell lysate.

      Lane 1 (input): SK-BR-3 whole cell lysate (10µg)

      Lane 2 (+): ab75852 + SK-BR-3 whole cell lysate (10µg).

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab75852 in SK-BR-3 whole cell lysate.

      For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      All lanes : Anti-Cathepsin D antibody [EPR3057Y] (ab75852) at 1/10000 dilution (unpurified)

      Lane 1 : MCF-7 cell lysate
      Lane 2 : A431 cell lysate
      Lane 3 : SKBR3 cell lysate
      Lane 4 : HepG2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size : 46 kDa
      Observed band size : 46 kDa
      Additional bands at : 28 kDa (possible cleavage fragment).      Bands at 28kDa are the Cathepsin D heavy chain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

    References for Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

    This product has been referenced in:
    • Sadleir KR  et al. Presynaptic dystrophic neurites surrounding amyloid plaques are sites of microtubule disruption, BACE1 elevation, and increased Aß generation in Alzheimer's disease. Acta Neuropathol 132:235-56 (2016). WB ; Mouse . Read more (PubMed: 26993139) »
    • Santaguida S  et al. Aneuploidy-induced cellular stresses limit autophagic degradation. Genes Dev 29:2010-21 (2015). Read more (PubMed: 26404941) »

    See all 4 Publications for this product

    Publishing research using ab75852? Please let us know so that we can cite the reference in this datasheet.

    Product Wall

    Submit an abreview for ab75852 Submit a question for ab75852

    6 Abreviews or Q&A

    10
    Votes: Highest First

    Western blot abreview for Anti-Cathepsin D antibody [EPR3057Y]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (blood monocyte derived macrophage)
    Loading amount 100 µg
    Specification blood monocyte derived macrophage
    Gel Running Conditions Reduced Denaturing (10)
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
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    Verified customer

    Submitted Jan 18 2013

    danke für die Informationen.
    Hier ein kurzes Update zu ihren Vorschlägen.

    1. HIER ist ein Verfahren, welches wir bei unseren Proben nicht anwenden können. Ich habe daher auch nicht das entsprechende Equipment. Werde jedoch v...

    Read More

    Vielen Dank für dieses Update.

    Es freut mich zu hören, dass Sie versuchen werden das Ergebnis mit unseren Vorschlägen zu verbessern. Ich weiß, dass es viel Zeit und Organisation im Labor erfordert und schätze Ihren E...

    Read More

    anbei das ausgefüllte Infoblatt mit angehängtem Protokoll.
    Haben sie schon etwas rausfinden können?

    Mit freundlichen Grüßen

    Vielen Dank für Ihren Anruf und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

    Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

    Das verwendete Protokoll sieht gut aus...

    Read More

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cathepsin D antibody [EPR3057Y]

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (liver)
    Specification liver
    Fixative Formaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH6.0
    Permeabilization No
    Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More
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    Verified customer

    Submitted Jun 11 2010

    Western blot abreview for Anti-Cathepsin D antibody [EPR3057Y]

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (MCF7 cells)
    Loading amount 15 µg
    Specification MCF7 cells
    Gel Running Conditions Reduced Denaturing (4-12%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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    Verified customer

    Submitted May 21 2009

    Western blot abreview for Anti-Cathepsin D antibody [EPR3057Y]

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (NIH 3T3 cells)
    Loading amount 15 µg
    Specification NIH 3T3 cells
    Gel Running Conditions Reduced Denaturing (4-12%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
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    Submitted May 21 2009

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"