使用敲除细胞株进行验证
重组
RabMAb

Anti-KSR1抗体[EPR2421Y] (ab68483)

概述

  • 产品名称
    Anti-KSR1抗体[EPR2421Y]
    参阅全部 KSR1 一抗
  • 描述
    兔单克隆抗体[EPR2421Y] to KSR1
  • 经测试应用
    适用于: WB, IHC-Pmore details
    不适用于: Flow Cyt,ICC or IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues near the C-terminus of human KSR.

  • 阳性对照
    • 293 cell lysate.
  • 常规说明

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab68483 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 - 1/2000. Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse and rat samples.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

  • 应用说明
    Is unsuitable for Flow Cyt,ICC or IP.
  • 靶标

    • 功能
      Location-regulated scaffolding protein connecting MEK to RAF. Promotes MEK and RAF phosphorylation and activity through assembly of an activated signaling complex. By itself, it has no demonstrated kinase activity.
    • 序列相似性
      Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
      Contains 1 phorbol-ester/DAG-type zinc finger.
      Contains 1 protein kinase domain.
    • 翻译后修饰
      Phosphorylated on Ser-309 and, to a higher extent, on Ser-404 by MARK3. Dephosphorylated on Ser-404 by PPP2CA. In resting cells, phosphorylated KSR1 is cytoplasmic and in stimulated cells, dephosphorylated KSR1 is membrane-associated.
    • 细胞定位
      Cytoplasm. Membrane. In unstimulated cells, where the phosphorylated form is bound to a 14-3-3 protein, sequestration in the cytoplasm occurs. Following growth factor treatment, the protein is free for membrane translocation, and it moves from the cytoplasm to the cell periphery.
    • Information by UniProt
    • 数据库链接
    • 别名
      • BKSR 1 antibody
      • BKSR1 antibody
      • dKsr antibody
      • EK 31 antibody
      • EK31 antibody
      • hb antibody
      • Kinase suppressor of Ras 1 antibody
      • Kinase suppressor of ras antibody
      • KSR 1 antibody
      • KSR antibody
      • KSR1 antibody
      • KSR1_HUMAN antibody
      • Positive Ras signaling mediator family member (86.4 kD) antibody
      • Positive Ras signaling mediator family member antibody
      • RSU 2 antibody
      • RSU2 antibody
      • SR 31 antibody
      • SR31 antibody
      • Suppressor of activated let 60 Ras SUR3 antibody
      • Suppressor of Ras1 31 antibody
      see all

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling KSR1 with Purified ab68483 at 1:100 dilution (1.49 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.



    • Predicted band size : 102 kDa

      Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: KSR1 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 - ab68483 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Unpurified  ab68483 was shown to recognize KSR1  when KSR1  knockout samples were used, along with additional cross-reactive bands. Wild-type and KSR1  knockout samples were subjected to SDS-PAGE. Ab68483 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1:10,000 dilution respectively and incubated overnight at 4C. 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.

    • Unpurified ab68483 staining KSR1 in NIH/3T3 (mouse embryo) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/230. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black)

      Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    • Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/1000 dilution (purified) + MEF (Mouse embryonic fibroblast (immortalized)) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 102 kDa
      Observed band size : 110 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST

    • Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/1000 dilution (purified) + 293 (Human embryonic kidney epithelial cell) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 102 kDa
      Observed band size : 110 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST

    • Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/1000 dilution (purified) + Rat-1 (Rat embryonic fibroblast) whole cell lysates

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 102 kDa
      Observed band size : 110 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST

    • Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/2000 dilution (unpurified) + 293 cell lysate at 10 µg

      Secondary
      goat anti rabbit HRP at 1/2000 dilution

      Predicted band size : 102 kDa
      Observed band size : 120 kDa (why is the actual band size different from the predicted?)

    文献

    ab68483 has not yet been referenced specifically in any publications.

    客户评价及客户问答

    There are currently no Customer reviews or Questions for ab68483.
    Please use the links above to contact us or submit feedback about this product.

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

    注册