製品の概要

  • 製品名
    Anti-CAD antibody [EP710Y]
    CAD 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP710Y] to CAD
  • 特異性
    ab40800 is specific to the N-terminus of CAD.
  • アプリケーション
    適用あり: ICC/IF, WB, IP, Flow Cytmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CAD aa 1-100 (N terminal).
    Database link: P27708

  • ポジティブ・コントロール
    • WB: HeLa and HEK293 cell lysate. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa cell lysate.
  • 特記事項

    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.

    Mouse, 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-CAD antibody (Alexa Fluor® 647) [EP710Y] (ab203699)
    Anti-CAD antibody (HRP) [EP710Y] (ab203701)
    Anti-CAD antibody (Alexa Fluor® 568) [EP710Y] (ab208050)
    Anti-CAD antibody (Phycoerythrin) [EP710Y] (ab208994)

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

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab40800 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF 1/100 - 1/300.
WB 1/1000. Predicted molecular weight: 243 kDa.
IP 1/30 - 1/50.
Flow Cyt 1/120.

ターゲット情報

  • 関連性
    Carbamoyl phosphate synthetase-aspartate carbamoyltransferase-dihydroorotase (CAD) is a multifunctional protein that initiates and regulates mammalian de novo pyrimidine biosynthesis. This trifunctional protein which is associated with the enzymatic activities of the first 3 enzymes in the 6-step pathway of pyrimidine biosynthesis is the rate-limiting step in the de novo pyrimidine synthetic pathway. Although most of the CAD protein in the cell is cytosolic, phosphorylation at threonine 456 localizes the protein to the nucleus. While MAPK and EGF phosphorylate CAD at threonine 456, MAPK and c-myc have been found to induce over-expression of CAD.
  • 細胞内局在
    Cytoplasmic and Nuclear
  • 参照データベース
  • 別名
    • Aspartate carbamoyltransferase antibody
    • Aspartate transcarbamylase antibody
    • CAD protein antibody
    • CAD trifunctional protein antibody
    • Carbamoyl phosphate synthetase 2 antibody
    • Carbamoyl-phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase antibody
    • CPSase ATCase DHOase antibody
    • Dihydroorotase antibody
    • Glutamine dependent carbamoyl phosphate synthase antibody
    • Multifunctional protein CAD antibody
    see all

画像

  • Anti-CAD antibody [EP710Y] (ab40800) at 1/2000 dilution (purified) + HeLa cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 243 kDa
    Observed band size : 250 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of HeLa cells with purified ab40800 at a working dilution of 1/300, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab40800 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling CAD (red) with ab40800 at a 1/120 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

  • ab40800 (purified) at 1/30 immunoprecipitating CAD in 10 μg HeLa (Lanes 1 and 2, observed at 250 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Anti-CAD antibody [EP710Y] (ab40800) at 1/1000 dilution (purified) + HeLa cell lysate at 1/20000 dilution

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 243 kDa
    Observed band size : 250 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab40800 (1/100), staining human CAD (N-term) in HeLa cells by Immunofluorescence.

  • Anti-CAD antibody [EP710Y] (ab40800) at 1/1000 dilution (unpurified) + HEK293 whole cell lysate (ab7902) at 10 µg

    Predicted band size : 243 kDa
    Observed band size : 250 kDa (why is the actual band size different from the predicted?)

参考文献

This product has been referenced in:
  • Sun W  et al. Interaction between von Hippel-Lindau Protein and Fatty Acid Synthase Modulates Hypoxia Target Gene Expression. Sci Rep 7:7190 (2017). Read more (PubMed: 28775317) »
  • Rabinovich S  et al. Diversion of aspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis. Nature 527:379-383 (2015). WB . Read more (PubMed: 26560030) »

See all 4 Publications for this product

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

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