Name: Anti-Cathepsin D antibody [EPR3057Y]
Brand: RabMAb
SKU: ab75852
Rating: 5 (11 reviews)

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Western blot - 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 (Intra), ICC/IF, WB, IP, IHC-P, IHC-Fr
Reacts with: Mouse, Human

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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 (Intra), ICC/IF, WB, IP, IHC-P, IHC-Frmore 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 (intra): HepG2 cells. IHC-Fr: Hu liver tissue sections.
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
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

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
- Neuroscience
- Diseases
- Neuroscience
- Processes

The Abpromise guarantee

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 (Intra)		Use at an assay dependent concentration.
ICC/IF	(1)	1/100 - 1/1000.
WB	(8)	1/2000 - 1/10000. Predicted molecular weight: 46 kDa.
IP		1/10 - 1/20.
IHC-P	(1)	1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
IHC-Fr	(1)	1/1000.

Notes
Flow Cyt (Intra) 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.
IHC-Fr 1/1000.

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

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

Cells were fixed with 4% paraformaldehyde and permeabilized 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.
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)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

ab75852 has been referenced in 72 publications.

Aoto K et al. ATP6V0A1 encoding the a1-subunit of the V0 domain of vacuolar H+-ATPases is essential for brain development in humans and mice. Nat Commun 12:2107 (2021). PubMed: 33833240
Mehrotra S et al. Expression of Cathepsins B, D, and G in Microcystic Lymphatic Malformation. Lymphat Res Biol 19:347-354 (2021). PubMed: 33337924
Lin J et al. H(+)/Cl(-) exchange transporter 7 promotes lysosomal acidification-mediated autophagy in mouse cardiomyocytes. Mol Med Rep 23:N/A (2021). PubMed: 33495814
Wang X et al. A small molecule binding HMGB1 inhibits caspase-11-mediated lethality in sepsis. Cell Death Dis 12:402 (2021). PubMed: 33854044
Sitia L et al. Selective Targeting of Cancer-Associated Fibroblasts by Engineered H-Ferritin Nanocages Loaded with Navitoclax. Cells 10:N/A (2021). PubMed: 33562504

View all Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application

Western blot

Sample

Mouse Tissue lysate - whole (brain)

Gel Running Conditions

Reduced Denaturing

Loading amount

40 µg

Specification

brain

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 07 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (brain section)

Permeabilization

Yes - 0.3% triton

Specification

brain section

Blocking step

Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 20°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Nov 13 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (kidney)

Gel Running Conditions

Reduced Denaturing (4-12% bis-tris gel, MES buffer)

Loading amount

30 µg

Specification

kidney

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 03 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (Dopaminergic Neurons)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

Dopaminergic Neurons

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 19 2020

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

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

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

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

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

1-10 of 11 Abreviews

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Assay kits

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (72)

Customer reviews and Q&As

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

Immunocytochemistry/ Immunofluorescence 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]

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

Immunohistochemistry (Frozen 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]

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

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

Post-translational
modifications