使用敲除细胞株进行验证RabMAb

Anti-Protein Kinase D2抗体[EP1495Y] (ab51250)

概述

  • 产品名称Anti-Protein Kinase D2抗体[EP1495Y]
    参阅全部 Protein Kinase D2 一抗
  • 描述
    兔单克隆抗体[EP1495Y] to Protein Kinase D2
  • 经测试应用适用于: WB, Flow Cyt, IHC-P, ICC/IFmore details
    不适用于: IP
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide (Human)

  • 阳性对照
    • HeLa cell lysate or human breast carcinoma tissue. IF/ICC: HeLa cell line
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

性能

应用

Our Abpromise guarantee covers the use of ab51250 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
WB 1/500. Detects a band of approximately 105 kDa (predicted molecular weight: 97 kDa).
Flow Cyt 1/20.

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

IHC-P Use at an assay dependent concentration.
ICC/IF 1/100 - 1/250.
  • 应用说明Is unsuitable for IP.
  • 靶标

    • 功能Converts transient diacylglycerol (DAG) signals into prolonged physiological effects, downstream of PKC. Involved in resistance to oxidative stress.
    • 组织特异性Widely expressed.
    • 序列相似性Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. PKD subfamily.
      Contains 1 PH domain.
      Contains 2 phorbol-ester/DAG-type zinc fingers.
      Contains 1 protein kinase domain.
    • 翻译后修饰Phosphorylation of Ser-876 correlates with the activation status of the kinase.
      Ser-706 is probably phosphorylated by PKC.
    • 细胞定位Cytoplasm. Membrane. Translocation to the cell membrane is required for kinase activation.
    • Information by UniProt
    • 数据库链接
    • 别名
      • DKFZp586E0820 antibody
      • HSPC187 antibody
      • KPCD2_HUMAN antibody
      • nPKC D2 antibody
      • nPKC-D2 antibody
      • PKD2 antibody
      • PRKD 2 antibody
      • Prkd2 antibody
      • Protein kinase D2 antibody
      • Serine/threonine protein kinase D2 antibody
      • Serine/threonine-protein kinase D2 antibody
      see all

    Anti-Protein Kinase D2 antibody [EP1495Y] 图像



    • Predicted band size : 97 kDa

      Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: Protein Kinase D2 knockout HAP1 cell lysate (40 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HEK293 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab51250 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab51250 was shown to recognize Protein Kinase D2 when Protein Kinase D2 knockout samples were used, along with additional cross-reactive bands. Wild-type and Protein Kinase D2 knockout samples were subjected to SDS-PAGE. Ab51250 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1:10,000 dilution respectively and incubated overnight at 4C. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.

    • Anti-Protein Kinase D2 antibody [EP1495Y] (ab51250) at 1/500 dilution + HeLa cell lysate at 10 µg

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

      Predicted band size : 97 kDa
      Observed band size : 105 kDa (why is the actual band size different from the predicted?)
    • Ab51250 (1:100) staining human Protein Kinase D2 in human breast carcinoma tissue by immunohistochemistry using paraffin embedded tissue.
    • ICC/IF image of ab51250 stained HeLa cells. The cells were 4% Formaldehyde fixed (10 mins) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51250, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    Anti-Protein Kinase D2 antibody [EP1495Y] (ab51250)参考文献

    This product has been referenced in:
    • Bhavanasi D  et al. Protein kinase Cd mediates the activation of protein kinase D2 in platelets. Biochem Pharmacol 82:720-7 (2011). WB ; Human, Mouse . Read more (PubMed: 21736870) »
    • Azoitei N  et al. Protein kinase D2 is a novel regulator of glioblastoma growth and tumor formation. Neuro Oncol 13:710-24 (2011). WB, IHC-P ; Human, Chicken . Read more (PubMed: 21727210) »

    See all 3 Publications for this product

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