Recombinant Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
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 (Intra), ICC/IF, WB, IP, IHC-P, IHC-Fr
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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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 (Intra), 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.
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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 (intra): HepG2 cells. IHC-Fr: Hu liver tissue sections.
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General notes
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped 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. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3057Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Cathepsin D antibody [EPR3057Y] (ab197605)
- Alexa Fluor® 647 Anti-Cathepsin D antibody [EPR3057Y] (ab198326)
- Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free (ab207549)
- Alexa Fluor® 594 Anti-Cathepsin D antibody [EPR3057Y] (ab207874)
- Alexa Fluor® 405 Anti-Cathepsin D antibody [EPR3057Y] (ab207875)
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
- Hep G2 whole cell lysate (ab166833)
- MCF7 whole cell lysate (ab29537)
- A-431 whole cell lysate (ab30132)
- Mouse brain tissue lysate - total protein (ab30151)
- 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)
- A-431 whole cell lysate (ab7909)
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
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ICC/IF | (1) |
1/100 - 1/1000.
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WB | (8) |
1/2000 - 1/10000. Predicted molecular weight: 46 kDa.
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IP |
1/10 - 1/20.
|
|
IHC-P | (1) |
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
IHC-Fr | (1) |
1/1000.
|
Notes |
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Flow Cyt (Intra)
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. |
IHC-Fr
1/1000. |
Target
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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
modificationsN- 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
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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
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Alternative names
- CatD antibody
- CATD_HUMAN antibody
- Cathepsin D antibody
see all
Images
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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.
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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.
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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).
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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.
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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.
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Intracellular Flow Cytometry analysis of HepG2 (Human liver hepatocellular carcinoma cell line)cells labeling Cathepsin D with purified ab75852 at 1/20 dilution (10µg/ml) (red).
Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 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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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.
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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) 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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (72)
ab75852 has been referenced in 72 publications.
- Aoto K et al. ATP6V0A1 encoding the a1-subunit of the V0 domain of vacuolar H+-ATPases is essential for brain development in humans and mice. Nat Commun 12:2107 (2021). PubMed: 33833240
- Mehrotra S et al. Expression of Cathepsins B, D, and G in Microcystic Lymphatic Malformation. Lymphat Res Biol 19:347-354 (2021). PubMed: 33337924
- Lin J et al. H(+)/Cl(-) exchange transporter 7 promotes lysosomal acidification-mediated autophagy in mouse cardiomyocytes. Mol Med Rep 23:N/A (2021). PubMed: 33495814
- Wang X et al. A small molecule binding HMGB1 inhibits caspase-11-mediated lethality in sepsis. Cell Death Dis 12:402 (2021). PubMed: 33854044
- Sitia L et al. Selective Targeting of Cancer-Associated Fibroblasts by Engineered H-Ferritin Nanocages Loaded with Navitoclax. Cells 10:N/A (2021). PubMed: 33562504