Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
- Datasheet
- Specific References (13)
- Protocols
Overview
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Product nameAnti-Cathepsin D antibody [EPR3057Y]
See all Cathepsin D primary antibodies -
DescriptionRabbit monoclonal [EPR3057Y] to Cathepsin D
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Host speciesRabbit
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Tested applicationsSuitable for: Flow Cyt, ICC/IF, WB, IP, IHC-Pmore details
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Species reactivityReacts with: Mouse, Human
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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).
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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.
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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.
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.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR3057Y
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
- Anti-Cathepsin D antibody [EPR3057Y] (Alexa Fluor® 488) (ab197605)
- Anti-Cathepsin D antibody [EPR3057Y] (Alexa Fluor® 647) (ab198326)
- Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free (ab207549)
- Anti-Cathepsin D antibody [EPR3057Y] (Alexa Fluor® 594) (ab207874)
- Anti-Cathepsin D antibody [EPR3057Y] (Alexa Fluor® 405) (ab207875)
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Assay kits
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Positive Controls
- HepG2 whole cell lysate (ab166833)
- MCF7 whole cell lysate (ab29537)
- A431 whole cell lysate (ab30132)
- Mouse brain tissue lysate - total protein (ab30151)
- MCF7 whole cell lysate (ab3871)
- Mouse brain tissue lysate - total protein (0 days) (ab7188)
- Mouse brain tissue lysate - total protein (14 days) (ab7189)
- Mouse brain tissue lysate - total protein (7 days) (ab7190)
- HepG2 whole cell lysate (ab7900)
- A431 whole cell lysate (ab7909)
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. |
Target
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FunctionAcid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
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Tissue specificityExpressed in the aorta extrcellular space (at protein level).
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Involvement in diseaseCeroid lipofuscinosis, neuronal, 10
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Sequence similaritiesBelongs to the peptidase A1 family.
Contains 1 peptidase A1 domain. -
Post-translational
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
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Database links
- Entrez Gene: 1509 Human
- Entrez Gene: 13033 Mouse
- Omim: 116840 Human
- SwissProt: P07339 Human
- SwissProt: P18242 Mouse
- Unigene: 654447 Human
- Unigene: 231395 Mouse
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Alternative names
- CatD antibody
- CATD_HUMAN antibody
- Cathepsin D antibody
see all
Images
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-7-ab75852IHC1.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-6-ab75852ICCIF1.jpg)
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).
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![Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-9-ab75852WB2.jpg)
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.
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![Flow Cytometry - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-273201-ab75852FC1.jpg)
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.
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![Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-8-ab75852WB1.jpg)
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
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 and dilution buffer: 5% NFDM /TBST.
ab75852 detects an extra band at 75kDa.
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![Immunoprecipitation - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-5-ab75852IP1.jpg)
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.
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![Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852wb.jpg)
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
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)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852.gif)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)](http://a.abcam.com/ps/products/75/ab75852/Images/ab75852-2.gif)
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.
Protocols
References
This product has been referenced in:
- Wang J et al. Silica nanoparticles induce autophagy dysfunction via lysosomal impairment and inhibition of autophagosome degradation in hepatocytes. Int J Nanomedicine 12:809-825 (2017). Read more (PubMed: 28182147) »
- Zahur M et al. Long-Term Assessment of AAV-Mediated Zinc Finger Nuclease Expression in the Mouse Brain. Front Mol Neurosci 10:142 (2017). Read more (PubMed: 28588449) »
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"