Name: Anti-Cathepsin D antibody [EPR3057Y]
Brand: RabMAb
SKU: ab75852
Rating: 5 (7 reviews)

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, ICC/IF, WB, IP, IHC-Pmore 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.
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.
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.
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.
This product is a recombinant rabbit monoclonal antibody.

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

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
Flow Cyt		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. See IHC antigen retrieval protocols.

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
modifications

N- 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).
Target information above from: UniProt accession P07339 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 1509 Human
- Entrez Gene: 13033 Mouse
- Omim: 116840 Human
- SwissProt: P07339 Human
- SwissProt: P18242 Mouse
- Unigene: 654447 Human
- Unigene: 231395 Mouse
Alternative names
- CatD antibody
- CATD_HUMAN antibody
- Cathepsin D antibody
- Cathepsin D heavy chain antibody
- CD antibody
- Ceroid lipofuscinosis neuronal 10 antibody
- CLN10 antibody
- CPSD antibody
- ctsd antibody
- Epididymis secretory sperm binding protein Li 130P antibody
- HEL S 130P antibody
- Lysosomal aspartyl peptidase antibody
- Lysosomal aspartyl protease antibody
- MGC2311 antibody
see all

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.
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, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10,000).

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.
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.

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Milenkovic A et al. BEST1 protein stability and degradation pathways differ between autosomal dominant Best disease and autosomal recessive bestrophinopathy accounting for the distinct retinal phenotypes. Hum Mol Genet 27:1630-1641 (2018). Read more (PubMed: 29668979) »
Härtlova A et al. LRRK2 is a negative regulator of Mycobacterium tuberculosis phagosome maturation in macrophages. EMBO J 37:N/A (2018). Read more (PubMed: 29789389) »

See all 22 Publications for this product

Publishing research using ab75852? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-7 of 7 Abreviews

Application

Western blot

Sample

Dog Cell lysate - whole cell (MDCK2 cells)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

20 µg

Treatment

50uM chloroquine for 5h

Specification

MDCK2 cells

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 24 2018

Abcam has not validated the combination of species/application used in this Abreview.

Application

Immunohistochemistry (Frozen sections)

Sample

Human Tissue sections (adult skin)

Permeabilization

Specification

adult skin

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C

Fixative

unfixed

Abcam user community

Verified customer

Submitted Jul 02 2018

Application

Western blot

Sample

Human Cell lysate - whole cell (Primary keratinocytes)

Gel Running Conditions

Reduced Denaturing (any kD BioRad)

Loading amount

50 µg

Specification

Primary keratinocytes

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Julien Coutier

Verified customer

Submitted Jul 02 2018

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (blood monocyte derived macrophage)

Loading amount

100 µg

Specification

blood monocyte derived macrophage

Gel Running Conditions

Reduced Denaturing (10)

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (liver)

Specification

liver

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate pH6.0

Permeabilization

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Jun 11 2010

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (MCF7 cells)

Loading amount

15 µg

Specification

MCF7 cells

Gel Running Conditions

Reduced Denaturing (4-12%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted May 21 2009

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Mouse Cell lysate - whole cell (NIH 3T3 cells)

Loading amount

15 µg

Specification

NIH 3T3 cells

Gel Running Conditions

Reduced Denaturing (4-12%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted May 21 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-Cathepsin D antibody [EPR3057Y] (ab75852)

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References

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Western blot abreview for Anti-Cathepsin D antibody [EPR3057Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cathepsin D antibody [EPR3057Y]

Western blot abreview for Anti-Cathepsin D antibody [EPR3057Y]

Western blot abreview for Anti-Cathepsin D antibody [EPR3057Y]