製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to VPS35
  • アプリケーション
    適用あり: WB, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat, Rabbit, Horse, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human VPS35.

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: HepG2; MOLT4; Raji. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
    Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab118838 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 91 kDa (predicted molecular weight: 91 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml.

ターゲット情報

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  • Predicted band size : 91 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
    Lane 3: NIH/3T3 cell lysate (20 µg)
    Lane 4: A549 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab118838 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab118838 was shown to recognize VPS35 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab118838 and ab8245 (loading control to GAPDH) were diluted 1μg/mL and 1/2000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • ICC/IF image of ab118838 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 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 ab118838 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

  • All lanes : Anti-VPS35 antibody (ab118838) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate
    Lane 3 : Raji (Human Burkitt's lymphoma 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 : 91 kDa
    Observed band size : 85 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 24 kDa,35 kDa,71 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

参考文献

ab118838 has not yet been referenced specifically in any publications.

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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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