Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3054] to proCathepsin D
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
参阅全部 proCathepsin D 一抗
描述兔单克隆抗体[EPR3054] to proCathepsin D
经测试应用适用于: WB, IHC-P, Flow Cyt, ICC/IFmore details
Synthetic peptide within Human proCathepsin D aa 1-100. The exact sequence is proprietary.
- Human breast ductal infiltrating carcinoma tissue; A431, MCF7 and SKBR3 cell lysates.
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.
存放说明Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
解离常数（KD）KD = 1.16 x 10 -10 M Learn more about KD
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
Concentration information loading...
纯度Protein A purified
Our Abpromise guarantee covers the use of ab134169 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 - 1/10000. Predicted molecular weight: 44 kDa.|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.|
功能Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
疾病相关Defects in CTSD are the cause of neuronal ceroid lipofuscinosis type 10 (CLN10) [MIM:610127]; also known as neuronal ceroid lipofuscinosis due to cathepsin D deficiency. A form of neuronal ceroid lipofuscinosis with onset at birth or early childhood. Neuronal ceroid lipofuscinoses are progressive neurodegenerative, lysosomal storage diseases characterized by intracellular accumulation of autofluorescent liposomal material, and clinically by seizures, dementia, visual loss, and/or cerebral atrophy.
序列相似性Belongs to the peptidase A1 family.
细胞定位Lysosome. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
- CATD_HUMAN antibody
- cathepsin D (lysosomal aspartyl protease) antibody
- Cathepsin D antibody
All lanes : Anti-proCathepsin D antibody [EPR3054] (ab134169) at 1/2000 dilution
Lane 1 : Wild-type A431 whole cell lysate
Lane 2 : Cathepsin D 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: 46 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green - ab134169 observed at 46 kDa. Red - loading control, ab7291 (mouse anti-tubulin), observed at 50 kDa.
ab134169 was shown to recognize in wild-type A431 cells as signal was lost at the expected MW in CTSD knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CTSD knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab134169 and ab7291 (Mouse anti-tubulin loading control) were incubated overnight at 4°C at 1/2000 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.
All lanes : Anti-proCathepsin D antibody [EPR3054] (ab134169) at 1/2000 dilution (purified)
Lane 1 : MCF-7 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : SKBR-3 cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 44 kDa
Observed band size: 44 kDa
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling proCathepsin D with purified ab134169 at 1/100 dilution (10ug/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.
Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab134169 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunofluorescence staining of MCF7 cells with purified ab134169 at a working dilution of 1 in 50, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti rabbit (ab150077), used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab134169 was used at a dilution of 1/200 followed by an Alexa Fluor® 594 goat anti-mouse antibody at a dilution of 1/500.
All lanes : Anti-proCathepsin D antibody [EPR3054] (ab134169) at 1/2000 dilution (unpurified)
Lane 1 : MCF7 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : SKBR3 cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa
Immunohistochemical analysis of paraffin-embedded Human breast ductal infiltrating carcinoma tissue, staining proCathepsin D using unpurified ab134169 at a 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD
ab134169 被引用在 1 文献中.
- 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