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使用敲除细胞株进行验证重组RabMAb

重组Anti-DEPDC5抗体[EPR20497-23] - BSA and Azide free (ab272399)

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Western blot - Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)
  • Western blot - Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)
  • Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20497-23] to DEPDC5 - BSA and Azide free
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Unconjugated

概述

  • 产品名称

    Anti-DEPDC5抗体[EPR20497-23] - BSA and Azide free
    参阅全部 DEPDC5 一抗

  • 描述

    兔单克隆抗体[EPR20497-23] to DEPDC5 - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WBmore details
    不适用于: IHC-P or IP

  • 种属反应性

    与反应: Mouse, Human
    不与反应: Rat

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Wild-type mouse E14 brain lysate. A549 cell lysate. Human Brain cell lysate

  • 常规说明

    ab272399 is the carrier-free version of ab213181.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab272399于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
Use at an assay dependent concentration. Detects a band of approximately 181 kDa (predicted molecular weight: 181 kDa).
说明
WB
Use at an assay dependent concentration. Detects a band of approximately 181 kDa (predicted molecular weight: 181 kDa).
应用说明
Is unsuitable for IHC-P or IP.

靶标

图片

  • Western blot - Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)
    Western blot - Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)
    All lanes : Anti-DEPDC5 antibody [EPR20497-23] (ab213181) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : DEPDC5 knockout A549 cell lysate
    Lane 3 : Human brain tissue lysate
    Lane 4 : Mouse brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 181 kDa
    Observed band size: 181 kDa



    False colour image of Western blot: Anti-DEPDC5 antibody [EPR20497-23] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab213181 was shown to bind specifically to DEPDC5. A band was observed at 181 kDa in wild-type A549 cell lysates with no signal observed at this size in DEPDC5 knockout cell line ab266906 (knockout cell lysate ab258394). To generate this image, wild-type and DEPDC5 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)
    Western blot - Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The wild-type and DEPDC5 knockout mouse E14 brain tissue lysates were kindly provided by an anonymous collaborator.

    ab213181 was shown to specifically react with DEPDC5 in wild-type mouse E14 brain tissue as signal was lost in DEPDC5 knockout tissue. Wild-type and DEPDC5 knockout samples were subjected to SDS-PAGE. ab213181 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100, 000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique. 

    Exposure time: 26 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, BSA and sodium azide (ab213181).

  • Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)
    Anti-DEPDC5 antibody [EPR20497-23] - BSA and Azide free (ab272399)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (0)

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