RabMAb

Anti-Cathepsin D抗体[EPR3057Y] (ab75852)

概述

  • 产品名称Anti-Cathepsin D抗体[EPR3057Y]
    参阅全部 Cathepsin D 一抗
  • 描述
    兔单克隆抗体[EPR3057Y] to Cathepsin D
  • 经测试应用适用于: ICC/IF, WB, IP, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cathepsin D aa 350 to the C-terminus (C terminal).

  • 阳性对照
    • WB: MCF-7, A431, SK-BR-3 and HepG2 cell lysates and mouse brain tissue lysate. IHC-P: Human breast carcinoma and liver tissues. ICC/IF: MCF-7 cells. IP: SK-BR-3 cell lysate.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Alternative versions available:

    Anti-Cathepsin D antibody (Alexa Fluor® 647) [EPR3057Y] (ab198326)

    Anti-Cathepsin D antibody (Alexa Fluor® 488) [EPR3057Y] (ab197605)

     

性能

应用

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.

应用 Ab评论 说明
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 protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

靶标

  • 功能Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
  • 组织特异性Expressed in the aorta extrcellular space (at protein level).
  • 疾病相关Ceroid lipofuscinosis, neuronal, 10
  • 序列相似性Belongs to the peptidase A1 family.
    Contains 1 peptidase A1 domain.
  • 翻译后修饰N- and O-glycosylated.
  • 细胞定位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
  • 数据库链接
  • 别名
    • CatD antibody
    • CATD_HUMAN 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

Anti-Cathepsin D antibody [EPR3057Y] 图像

  • 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 and dilution buffer: 5% NFDM /TBST.

  • All lanes : Anti-Cathepsin D antibody [EPR3057Y] (ab75852) at 1/5000 dilution (purified)

    Lane 1 : MCF-7 whole cell lysate
    Lane 2 : SKBR-3 whole cell lysate
    Lane 3 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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 and dilution buffer: 5% NFDM /TBST.

    ab75852 detects an extra band at 75kDa.

  • 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.

  • 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).

  • 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, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-Cathepsin D antibody [EPR3057Y] (ab75852) at 1/10000 dilution (unpurified)

    Lane 1 : MCF-7 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : SKBR3 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    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) analysis of human breast carcinoma tissue labelling Cathepsin D with unpurified ab75852 at a dilution of 1/500.

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

Anti-Cathepsin D antibody [EPR3057Y] (ab75852)参考文献

This product has been referenced in:
  • Sadleir KR  et al. Presynaptic dystrophic neurites surrounding amyloid plaques are sites of microtubule disruption, BACE1 elevation, and increased Aß generation in Alzheimer's disease. Acta Neuropathol 132:235-56 (2016). WB ; Mouse . Read more (PubMed: 26993139) »
  • Santaguida S  et al. Aneuploidy-induced cellular stresses limit autophagic degradation. Genes Dev 29:2010-21 (2015). Read more (PubMed: 26404941) »

See all 4 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 Jan 18 2013

Vielen Dank für dieses Update.

Es freut mich zu hören, dass Sie versuchen werden das Ergebnis mit unseren Vorschlägen zu verbessern. Ich weiß, dass es viel Zeit und Organisation im Labor erfordert und schätze Ihren E...

Read More

Vielen Dank für Ihren Anruf und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

Das verwendete Protokoll sieht gut aus...

Read More
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 No
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 May 21 2009

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