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    products/primary-antibodies/ksr1-antibody-epr2421y-ab68483.pdf

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Signal Transduction Protein Phosphorylation Tyrosine Kinases Other
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-KSR1 antibody [EPR2421Y] (ab68483)

  • Datasheet
  • SDS
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Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KSR1 antibody [EPR2421Y] (ab68483)
  • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
  • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
  • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
  • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
  • Anti-KSR1 antibody [EPR2421Y] (ab68483)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2421Y] to KSR1
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-KSR1 antibody [EPR2421Y]
    See all KSR1 primary antibodies
  • Description

    Rabbit monoclonal [EPR2421Y] to KSR1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: Wild-type HAP1 cell lysate. HeLa, MEF, Rat1 and HEK-293 cell lysate. IHC-P: Human cervical carcinoma tissue.
  • General notes

    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.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2421Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Other
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases

Associated products

  • Alternative Versions

    • Anti-KSR1 antibody [EPR2421Y] - BSA and Azide free (ab227109)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human KSR1 protein (ab190396)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
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 IHC antigen retrieval protocols.

Notes
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 IHC antigen retrieval protocols.

Application notes
Is unsuitable for Flow Cyt,ICC/IF or IP.

Target

  • Function

    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.
  • Sequence similarities

    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.
  • Post-translational
    modifications

    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.
  • Cellular localization

    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.
  • Target information above from: UniProt accession Q8IVT5 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8844 Human
    • Entrez Gene: 16706 Mouse
    • Entrez Gene: 360573 Rat
    • Omim: 601132 Human
    • SwissProt: Q8IVT5 Human
    • SwissProt: Q61097 Mouse
    • Unigene: 133534 Human
    • Unigene: 4745 Mouse
    • Alternative names

      • 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

    Images

    • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KSR1 antibody [EPR2421Y] (ab68483)

      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.

    • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      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

    • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/1000 dilution (purified) + HEK-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

    • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/1000 dilution (purified) + Rat1 (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

    • Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Western blot - Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Anti-KSR1 antibody [EPR2421Y] (ab68483) at 1/2000 dilution (unpurified) + HEK-293 (Human epithelial cell line from embryonic kidney) 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?

    • Anti-KSR1 antibody [EPR2421Y] (ab68483)
      Anti-KSR1 antibody [EPR2421Y] (ab68483)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (3)

    Publishing research using ab68483? Please let us know so that we can cite the reference in this datasheet.

    ab68483 has been referenced in 3 publications.

    • Rao C  et al. KSR1- and ERK-dependent translational regulation of the epithelial-to-mesenchymal transition. Elife 10:N/A (2021). PubMed: 33970103
    • Bai Y  et al. Effect of lentiviral vector-mediated KSR1 gene silencing on the proliferation of renal tubular epithelial cells and expression of inflammatory factors in a rat model of ischemia/reperfusion injury. Acta Biochim Biophys Sin (Shanghai) 50:807-816 (2018). PubMed: 30020400
    • Lavoie H  et al. MEK drives BRAF activation through allosteric control of KSR proteins. Nature 554:549-553 (2018). PubMed: 29433126

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