Recombinant Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free (ab207549)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-Fr, ICC/IF, IP, IHC-P
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free
See all Cathepsin D primary antibodies -
Description
Rabbit monoclonal [EPR3057Y] to Cathepsin D - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Fr, ICC/IF, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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
ab207549 is the carrier-free version of ab75852.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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)
- Alexa Fluor® 594 Anti-Cathepsin D antibody [EPR3057Y] (ab207874)
- Alexa Fluor® 405 Anti-Cathepsin D antibody [EPR3057Y] (ab207875)
- PE Anti-Cathepsin D antibody [EPR3057Y] (ab305650)
- APC Anti-Cathepsin D antibody [EPR3057Y] (ab305651)
- HRP Anti-Cathepsin D antibody [EPR3057Y] (ab305652)
- Alexa Fluor® 555 Anti-Cathepsin D antibody [EPR3057Y] (ab312170)
- Alexa Fluor® 568 Anti-Cathepsin D antibody [EPR3057Y] (ab312657)
- Anti-Cathepsin D antibody [EPR3057Y] (ab75852)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab207549 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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WB |
Use at an assay dependent concentration. Detects a band of approximately 28, 46 kDa (predicted molecular weight: 44 kDa).
Please check the parent abID, ab75882, for more information on dilutions. |
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IHC-Fr |
1/1000.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 28, 46 kDa (predicted molecular weight: 44 kDa). Please check the parent abID, ab75882, for more information on dilutions. |
IHC-Fr
1/1000. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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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Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free (ab207549) + SK-BR-3 (human mammary gland adenocarcinoma) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Predicted band size: 44 kDa
Observed band size: 28, 46 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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This data was developed using the same antibody clone in a different buffer formulation (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.
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Intracellular 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 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).
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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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Cathepsin D with unpurified ab75852 at a dilution of 1/500.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (3)
ab207549 has been referenced in 3 publications.
- Martinelli S et al. Stress-primed secretory autophagy promotes extracellular BDNF maturation by enhancing MMP9 secretion. Nat Commun 12:4643 (2021). PubMed: 34330919
- Slesiona S et al. Persistence versus escape: Aspergillus terreus and Aspergillus fumigatus employ different strategies during interactions with macrophages. PLoS One 7:e31223 (2012). ICC/IF . PubMed: 22319619
- Xie LQ et al. Novel proteomic strategy reveal combined alpha1 antitrypsin and cathepsin D as biomarkers for colorectal cancer early screening. J Proteome Res 9:4701-9 (2010). WB, IHC-P ; Human . PubMed: 20666480