Recombinant
RabMAb

Recombinant Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free (ab207549)

Overview

  • Product name

    Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free
    See all Cathepsin D primary antibodies
  • Description

    Rabbit monoclonal [EPR3057Y] to Cathepsin D - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, 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

    • MCF-7, A431, SKBR3 and HepG2 cell lysates. Human breast carcinoma tissue. Human liver tissue.
  • General notes

    Ab207549 is the carrier-free version of ab75852. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab207549 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab207549 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
WB Use at an assay dependent concentration. Detects a band of approximately 28, 46 kDa (predicted molecular weight: 44 kDa).

Please check the parent abID, ab75882, for more information on dilutions.

Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • 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).
  • Information by UniProt
  • Database links

  • 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

Images

  • Anti-Cathepsin D antibody [EPR3057Y] - BSA and Azide free (ab207549) + SK-BR-3 (human mammary gland adenocarcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 44 kDa
    Observed band size: 28, 46 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling Cathepsin D with purified ab75852 at 1/20 dilution (10µg/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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Cathepsin D with purified ab75852 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).

  • ab75852 (purified) at 1/20 immunoprecipitating Cathepsin D in SK-BR-3 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 buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75852).

References

This product has been referenced in:

  • Slesiona S  et al. Persistence versus escape: Aspergillus terreus and Aspergillus fumigatus employ different strategies during interactions with macrophages. PLoS One 7:e31223 (2012). ICC/IF . Read more (PubMed: 22319619) »
  • Xie LQ  et al. Novel proteomic strategy reveal combined alpha1 antitrypsin and cathepsin D as biomarkers for colorectal cancer early screening. J Proteome Res 9:4701-9 (2010). WB, IHC-P ; Human . Read more (PubMed: 20666480) »
See all 2 Publications for this product

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