Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3056Y] to Cathepsin D
- Suitable for: Flow Cyt, WB, IHC-P, ICC
- Knockout validated
- Reacts with: Human
Product nameAnti-Cathepsin D antibody [EPR3056Y]
See all Cathepsin D primary antibodies
DescriptionRabbit monoclonal [EPR3056Y] to Cathepsin D
Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ICCmore details
Species reactivityReacts with: Human
Synthetic peptide within Human Cathepsin D aa 50-150 (N terminal). The exact sequence is proprietary.
- WB: A431, HepG2 and MCF7 whole cell lysate. MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate. IHC-P: Human thyroid cancer tissue. ICC: MCF-7 cells. Flow Cyt: MCF-7 cells.
Mouse, 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
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS
Concentration information loading...
PurityProtein A purified
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.
|WB||1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
For unpurified use at 1/1000 - 1/2000.
|IHC-P||1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
For unpurified use at 1/500.
For unpurified use at 1/100 - 1/250.
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.
modificationsN- 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
- CatD antibody
- CATD_HUMAN antibody
- Cathepsin D antibody
Anti-Cathepsin D antibody [EPR3056Y] (ab75811) at 1/1000 dilution + MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 14,44 kDa why is the actual band size different from the predicted?
All lanes : Anti-Cathepsin D antibody [EPR3056Y] (ab75811) at 1/1000 dilution (Unpurified)
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 55 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.
Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cathepsin D with purified ab75811 at 1/50 dilution (1.88 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cathepsin D with purified ab75811 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid cancer tissue sections labeling Cathepsin D with purified ab75811 at 1/100 dilution (0.94 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
ab75811 has been referenced in 1 publication.
- Ashraf Y et al. Immunotherapy of triple-negative breast cancer with cathepsin D-targeting antibodies. J Immunother Cancer 7:29 (2019). PubMed: 30717773