Recombinant
RabMAb

Recombinant Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

Reviews (7)Q&A (2)References (45)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3057Y] to Cathepsin D
  • Suitable for: Flow Cyt, ICC/IF, WB, IP, IHC-P, IHC-Fr
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Cathepsin D antibody [EPR3057Y]
    See all Cathepsin D primary antibodies

  • Description

    Rabbit monoclonal [EPR3057Y] to Cathepsin D

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IP, IHC-P, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human Cathepsin D aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: MCF7, A431, SK-BR-3 and HepG2 whole cell lysate (ab7900) and mouse brain tissue lysate. IHC-P: Human breast carcinoma and liver tissues. ICC/IF: MCF7 cells. IP: SK-BR-3 cell lysate. Flow Cyt: HepG2 cells. IHC-Fr: Hu liver tissue sections.

  • General notes

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

    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.

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

    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.

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 IHC antigen retrieval protocols.
IHC-Fr 1/1000.

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

    • 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 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
      Lane 2 : SK-BR-3 (Human mammary gland adenocarcinoma cell line) whole cell lysate
      Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : 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/Dilution buffer: 5% NFDM /TBST.

       

    • Immunohistochemistry (Frozen sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
      Immunohistochemistry (Frozen sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

      IHC image of Cathepsin D staining in a section of frozen normal human liver performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab75852, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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

      Immunocytochemistry/Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling 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).

    • 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 labeling Cathepsin D with purified ab75852 at a dilution of 1/100.

      Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an 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.

    • 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/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 liver hepatocellular carcinoma cell line) cells labeling Cathepsin D with purified ab75852 at 1/20 dilution (10 µg/ml) (red).

      Cells were fixed with 4% paraformaldehyde and permeabilized 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.

    • 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 (Human mammary gland adenocarcinoma cell line) 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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

      Blocking/Dilution 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 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
      Lane 2 : A431 (Human epidermoid carcinoma cell line) cell lysate
      Lane 3 : SK-BR-3 (Human mammary gland adenocarcinoma cell line) cell lysate
      Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : 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 labeling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • 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 labeling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References

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

    ab75852 has been referenced in 45 publications.

    • Yang SY  et al. Glucocerebrosidase activity, cathepsin D and monomeric a-synuclein interactions in a stem cell derived neuronal model of a PD associated GBA1 mutation. Neurobiol Dis 134:104620 (2020). PubMed: 31634558
    • Ha Y  et al. Induction of Lysosome-associated Protein Transmembrane 4 Beta via Sulfatase 2 Enhances Autophagic Flux in Liver Cancer Cells. Hepatol Commun 3:1520-1543 (2019). PubMed: 31701075
    • Li H  et al. Mitochondrial dysfunction and mitophagy defect triggered by heterozygous GBA mutations. Autophagy 15:113-130 (2019). PubMed: 30160596
    • Su WF  et al. Overexpression of P2X4 receptor in Schwann cells promotes motor and sensory functional recovery and remyelination via BDNF secretion after nerve injury. Glia 67:78-90 (2019). PubMed: 30306657
    • He R  et al. TFEB-driven autophagy potentiates TGF-ß induced migration in pancreatic cancer cells. J Exp Clin Cancer Res 38:340 (2019). PubMed: 31387632
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Application

    Filter by Species

    Filter by Ratings

    1-7 of 7 Abreviews

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

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (MDCK2 cells)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    20 µg
    Treatment
    50uM chloroquine for 5h
    Specification
    MDCK2 cells
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Herr Dr. Vladimir Milenkovic

    Verified customer

    Submitted Oct 24 2018

    Immunohistochemistry (Frozen sections) abreview for Anti-Cathepsin D antibody [EPR3057Y]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (adult skin)
    Permeabilization
    No
    Specification
    adult skin
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    unfixed
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 02 2018

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

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Primary keratinocytes)
    Gel Running Conditions
    Reduced Denaturing (any kD BioRad)
    Loading amount
    50 µg
    Specification
    Primary keratinocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Mr. Julien Coutier

    Verified customer

    Submitted Jul 02 2018

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

     Good
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    Submitted Jan 18 2013

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

     Excellent
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    Submitted Jun 11 2010

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

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted May 21 2009

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

     Excellent
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    Submitted May 21 2009

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com