Recombinant Anti-proCathepsin D antibody [EPR3054] (ab134169)
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: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-proCathepsin D antibody [EPR3054]
See all proCathepsin D primary antibodies -
Description
Rabbit monoclonal [EPR3054] to proCathepsin D -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human proCathepsin D aa 1-100. The exact sequence is proprietary.
-
Positive control
- Human breast ductal infiltrating carcinoma tissue; A431, MCF7 and SKBR3 cell lysates.
-
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 1.16 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3054 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Assay kits
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
---|---|---|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
|
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.
|
|
ICC/IF |
1/100.
|
Notes |
---|
Flow Cyt (Intra)
Use at an assay dependent concentration. |
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. |
ICC/IF
1/100. |
Target
-
Function
Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease. -
Involvement in disease
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. -
Sequence similarities
Belongs to the peptidase A1 family. -
Cellular localization
Lysosome. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
-
Database links
- Entrez Gene: 1509 Human
- Omim: 116840 Human
- SwissProt: P07339 Human
- Unigene: 654447 Human
-
Alternative names
- CATD_HUMAN antibody
- cathepsin D (lysosomal aspartyl protease) antibody
- Cathepsin D antibody
see all
Images
-
All lanes : Anti-proCathepsin D antibody [EPR3054] (ab134169) at 1/2000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : Cathepsin D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
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.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 44 kDa
Observed band size: 44 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
-
Intracellular 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.
Secondary
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.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (1)
ab134169 has been referenced in 1 publication.
- 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