概述

  • 产品名称Anti-IDH2抗体
    参阅全部 IDH2 一抗
  • 描述
    兔多克隆抗体to IDH2
  • 经测试应用适用于: WB, ICC/IFmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Cow, Pig
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human IDH2.

  • 阳性对照
    • This antibody gave a positive signal in Human Heart, Skeletal Muscle, Liver and Brain Tissue Lysates, and in HepG2, DU 145 and SHSY5Y whole cell lysates.

性能

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应用

Our Abpromise guarantee covers the use of ab94359 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 Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 51 kDa).
ICC/IF Use a concentration of 1 µg/ml.

靶标

Anti-IDH2 antibody 图像



  • Predicted band size : 51 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: IDH2 knockout HAP1 cell lysate (20 µg)
    Lane 3: K562 cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab94359 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab94359 was shown to recognize IDH2 when IDH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and IDH2 knockout samples were subjected to SDS-PAGE. ab94359 at a concentration of 1ug/ml and ab8245 (loading control to GAPDH) diluted to 1/2000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-IDH2 antibody (ab94359) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 3 : Human liver tissue lysate - total protein (ab29889)
    Lane 4 : Human brain tissue lysate - total protein (ab29466)
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 51 kDa
    Observed band size : 47 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
    The predicted molecular weight of the IDH2 protein is 51-kDa. However, the protein sequence contains a 39 residue transit peptide at the N-terminus. This might explain the migration of the protein at a lower molecular weight than expected.
  • ICC/IF image of ab94359 stained HepG2 cells. The cells were 100% methanol fixed (5 min) 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 (ab94359, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% methanol fixed (5 min) MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml.

Anti-IDH2 antibody (ab94359)参考文献

ab94359 has not yet been referenced specifically in any publications.

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