Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-KSR1 antibody [EPR2421Y] - BSA and Azide free (ab227109)

Overview

  • Product name

    Anti-KSR1 antibody [EPR2421Y] - BSA and Azide free
    See all KSR1 primary antibodies
  • Description

    Rabbit monoclonal [EPR2421Y] to KSR1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • 293 cell lysate.
  • General notes

    Ab227109 is the carrier-free version of ab68483. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab227109 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab227109 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
IHC-P Use at an assay dependent concentration. 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.

WB Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa).
  • Application notes
    Is unsuitable for Flow Cyt,ICC 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.
    • Information by UniProt
    • Database links

    • 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

    • This WB data was generated using the same anti-KSR1 antibody clone, EPR2421Y, in a different buffer formulation (cat# 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.

    • 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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68483).

    • This IHC data was generated using the same anti-KSR1 antibody clone, EPR2421Y, in a different buffer formulation (cat# 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.

    References

    ab227109 has not yet been referenced specifically in any publications.

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