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リコンビナントRabMAb

Anti-CAD 抗体 [EP710Y] (ab40800)

Submit a review Submit a question References (9)
Western blot - Anti-CAD antibody [EP710Y] (ab40800)
  • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
  • Immunocytochemistry/ Immunofluorescence - Anti-CAD antibody [EP710Y] (ab40800)
  • Flow Cytometry (Intracellular) - Anti-CAD antibody [EP710Y] (ab40800)
  • Immunoprecipitation - Anti-CAD antibody [EP710Y] (ab40800)
  • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
  • Immunocytochemistry/ Immunofluorescence - Anti-CAD antibody [EP710Y] (ab40800)
  • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
  • Anti-CAD antibody [EP710Y] (ab40800)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP710Y] to CAD
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free PE

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

  • 由来種

    Rabbit

  • 特異性

    ab40800 is specific to the N-terminus of CAD.

  • アプリケーション

    適用あり: Flow Cyt (Intra), ICC/IF, WB, IPmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa and HEK293 cell lysate. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate (ab150035).

  • 特記事項

    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.

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

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab40800の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

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

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

    • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      Lanes 1 & 3 : Anti-CAD antibody [EP710Y] (ab40800) at 1/1000 dilution
      Lanes 2 & 4 : Anti-CAD antibody [EP711Y] (ab40765) at 1/1000 dilution

      Lanes 1-2 : NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate
      Lanes 3-4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


      Exposure time: 10 seconds


      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      ab40765 works better than ab40800 in western blot testing.

      We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using ab955 in western blot.

    • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      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

    • Immunocytochemistry/ Immunofluorescence - Anti-CAD antibody [EP710Y] (ab40800)
      Immunocytochemistry/ Immunofluorescence - Anti-CAD antibody [EP710Y] (ab40800)

      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 (Intracellular) - Anti-CAD antibody [EP710Y] (ab40800)
      Flow Cytometry (Intracellular) - Anti-CAD antibody [EP710Y] (ab40800)

      Intracellular 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 Fluorr® 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. 

       

       

    • Immunoprecipitation - Anti-CAD antibody [EP710Y] (ab40800)
      Immunoprecipitation - Anti-CAD antibody [EP710Y] (ab40800)

      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 for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

    • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      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

    • Immunocytochemistry/ Immunofluorescence - Anti-CAD antibody [EP710Y] (ab40800)
      Immunocytochemistry/ Immunofluorescence - Anti-CAD antibody [EP710Y] (ab40800)

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

    • Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      Western blot - Anti-CAD antibody [EP710Y] (ab40800)
      Anti-CAD antibody [EP710Y] (ab40800) at 1/1000 dilution (unpurified) + HEK-293 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?

    • Anti-CAD antibody [EP710Y] (ab40800)
      Anti-CAD antibody [EP710Y] (ab40800)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (9)

    ab40800 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab40800 は 9 報の論文で使用されています。

    • Mehazri L  et al. Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells. Int J Mol Sci 22:N/A (2021). PubMed: 33806191
    • Verrier ER  et al. Combined small molecule and loss-of-function screen uncovers estrogen receptor alpha and CAD as host factors for HDV infection and antiviral targets. Gut 69:158-167 (2020). WB ; Human . PubMed: 30833451
    • Brandt J  et al. The Cellular Protein CAD is Recruited into Ebola Virus Inclusion Bodies by the Nucleoprotein NP to Facilitate Genome Replication and Transcription. Cells 9:N/A (2020). PubMed: 32370067
    • Pesini A  et al. Brain pyrimidine nucleotide synthesis and Alzheimer disease. Aging (Albany NY) 11:8433-8462 (2019). PubMed: 31560653
    • Lee JS  et al. Urea Cycle Dysregulation Generates Clinically Relevant Genomic and Biochemical Signatures. Cell 174:1559-1570.e22 (2018). PubMed: 30100185
    • Sun W  et al. Interaction between von Hippel-Lindau Protein and Fatty Acid Synthase Modulates Hypoxia Target Gene Expression. Sci Rep 7:7190 (2017). PubMed: 28775317
    • Rabinovich S  et al. Diversion of aspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis. Nature 527:379-383 (2015). WB . PubMed: 26560030
    • DeVito SR  et al. Cytomegalovirus-mediated activation of pyrimidine biosynthesis drives UDP-sugar synthesis to support viral protein glycosylation. Proc Natl Acad Sci U S A 111:18019-24 (2014). PubMed: 25472841
    • Alfonso-Pérez T  et al. Cytoplasmic interaction of the tumour suppressor protein hSNF5 with dynamin-2 controls endocytosis. Oncogene N/A:N/A (2013). PubMed: 23851497

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