概述

  • 产品名称Anti-ARF6抗体
    参阅全部 ARF6 一抗
  • 描述
    兔多克隆抗体to ARF6
  • 经测试应用适用于: ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Chicken, Cow, Pig, Xenopus laevis
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus ARF6.

    (Peptide available as ab86625.)

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa whole cell, HEK293 whole cell, HepG2 whole cell, Human Liver Tissue

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab77581 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
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 20 kDa).

靶标

Anti-ARF6 antibody 图像



  • Predicted band size : 20 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ARF6 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab77581 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab77581 was shown to recognize ARF6 when ARF6 knockout samples were used, along with additional cross-reactive bands. Wild-type and ARF6 knockout samples were subjected to SDS-PAGE. ab77581 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-ARF6 antibody (ab77581) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 20 kDa
    Observed band size : 18 kDa (why is the actual band size different from the predicted?)
    We hypothesize that the 18 kDa band represents the mature form of ARF6.
  • ICC/IF image of ab77581 stained HeLa cells. The cells were 4% PFA 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 (ab77581, 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) HeLa cells at 1µg/ml.
  • IHC image of ARF6 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab77581, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-ARF6 antibody (ab77581) at 1 µg/ml

    Lane 1 : Lysates prepared from Mouse liver tissue.
    Lane 2 : Lysates prepared from Mouse lung tissue.
    Lane 3 : Lysates prepared from Rat liver tissue.
    Lane 4 : Lysates prepared from Rat lung tissue.

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated Pig polyclonal to rabbit IgG at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 20 kDa
    Observed band size : 21 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is a courtesy of Antibody Solutions Ltd

    See Abreview

Anti-ARF6 antibody (ab77581)参考文献

This product has been referenced in:

See all 5 Publications for this product

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Merci pour votre requête.

Le laboratoire a testé cet anticorps sur en gel migré BisTris 10% ou 4-12%, en tampon MES. La membrane était bloqué ensuite en 5%BSA en 0.1TBST. L'incubation de l'anticorps s'est fai...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Ms Liver, Ms Lung, Rat Liver, Rat Lung)
Loading amount 20 µg
Specification Ms Liver, Ms Lung, Rat Liver, Rat Lung
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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提交于 Jan 12 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"