Antibodies, Proteins, Kits and Reagents for Life Science

345 091 ₸ Product size

100 µl 345 091 ₸

Order now and get it on Tuesday March 02, 2021

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, ICC/IF, WB, IP, IHC-P, IHC-Fr
Reacts with: Mouse, Human

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 & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-Fr	Human
IHC-P	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human Cathepsin D aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.
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: HepG2 cells. IHC-Fr: Hu liver tissue sections.
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.
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
For more information see here.
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR3057Y
Isotype

IgG
Research areas

Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

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.
Immunohistochemistry (Frozen sections) - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

IHC image of Cathepsin D staining in a section of frozen normal human liver performed on a Leica BOND^TM 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.
Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

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).
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.
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/Dilution buffer: 5% NFDM /TBST.
Flow Cytometry - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

Flow Cytometry analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Cathepsin D with purified ab75852 at 1/20 dilution (10 µg/ml) (red).

Cells were fixed with 4% paraformaldehyde and permeabilized 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.
Immunoprecipitation - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

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.
Western blot - Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

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.
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.
Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

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