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RabMAb

Anti-Cdk4 抗体 [EPR2513Y] (ab68266)

製品の概要

  • 製品名
    Anti-Cdk4 antibody [EPR2513Y]
    Cdk4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR2513Y] to Cdk4
  • アプリケーション
    適用あり: ICC/IF, WB, IP, Flow Cytmore details
    適用なし: IHC-P
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues on the N-terminus of human CDK4.

  • ポジティブ・コントロール
    • WB: HeLa, MCF-7, Ramos and K562 cell lysates. ICC/IF: Wildtype HAP1 cells.
  • 特記事項

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

    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.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab68266 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 1/250.
WB 1/500 - 1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IP 1/30.
Flow Cyt 1/60.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • 追加情報
    Is unsuitable for IHC-P.
  • ターゲット情報

    • 機能
      Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.
    • 関連疾患
      Defects in CDK4 are a cause of susceptibility to cutaneous malignant melanoma type 3 (CMM3) [MIM:609048]. Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.
    • 配列類似性
      Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
      Contains 1 protein kinase domain.
    • 翻訳後修飾
      Phosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B.
    • 細胞内局在
      Cytoplasm. Nucleus. Membrane. Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G(1) phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G(1) to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus.
    • Information by UniProt
    • 参照データベース
    • 別名
      • Cdk 4 antibody
      • cdk4 antibody
      • CDK4 protein antibody
      • CDK4_HUMAN antibody
      • Cell division kinase 4 antibody
      • Cell division protein kinase 4 antibody
      • CMM 3 antibody
      • CMM3 antibody
      • Crk3 antibody
      • Cyclin dependent kinase 4 antibody
      • Cyclin-dependent kinase 4 antibody
      • Melanoma cutaneous malignant 3 antibody
      • MGC14458 antibody
      • p34 cdk4 antibody
      • PSK J3 antibody
      • PSK-J3 antibody
      see all

    画像



    • Predicted band size : 34 kDa

      Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 4 and 6: CDK4 knockout HAP1 cell lysate (20 µg)
      Lanes 1 and 2: Green signal from target - ab68266 observed at 34 kDa
      Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
      Lanes 5 and 6: Merged (red and green) signal
      ab68266 was shown to specifically react with CDK4 when CDK4 knockout samples were used. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab68266 and ab8226 (loading control to beta actin) were both diluted at 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • ab68266 staining Cdk4 in wild-type HAP1 cells (top panel) and Cdk4 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab68266 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • All lanes : Anti-Cdk4 antibody [EPR2513Y] (ab68266) at 1/1000 dilution

      Lane 1 : HeLa whole cell lysate
      Lane 2 : Mouse lung tissue lysate
      Lane 3 : Rat lung tissue lysate
      Lane 4 : Mouse spleen tissue lysate
      Lane 5 : Rat spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 34 kDa
      Observed band size : 34 kDa

      Lanes 1 - 5: Merged signal (red and green). Green - ab68266 observed at 34 kDa. Red - loading control, ab18058, observed at 130 kDa.

      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab68266 and ab18058 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

    • All lanes : Anti-Cdk4 antibody [EPR2513Y] (ab68266) at 1/500 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : Romas cell lysate
      Lane 4 : K452 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP conjugated Goat anti-rabbit at 1/2000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa

    参考文献

    This product has been referenced in:
    • Karst AM  et al. Modeling high-grade serous ovarian carcinogenesis from the fallopian tube. Proc Natl Acad Sci U S A 108:7547-52 (2011). Read more (PubMed: 21502498) »

    See 1 Publication for this product

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