Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-RIP antibody [EPR4689-100] (ab178420)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4689-100] to RIP
  • Suitable for: WB, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-RIP antibody [EPR4689-100]
    See all RIP primary antibodies

  • Description

    Rabbit monoclonal [EPR4689-100] to RIP

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cytmore details
    Unsuitable for: ICC,IHC-P or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human RIP. The exact sequence is proprietary.
    Database link: Q13546

  • Positive control

    • HeLa cells and cell lysates; Raji cell lysates.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab178420 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 1/1000 - 1/10000. Predicted molecular weight: 75 kDa.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
  • Application notes
    Is unsuitable for ICC,IHC-P or IP.

    • Target

    • Function

      Essential adapter molecule for the activation of NF-kappa-B. Following different upstream signals (binding of inflammatory cytokines, stimulation of pathogen recognition receptors, or DNA damage), particular RIPK1-containing complexes are formed, initiating a limited number of cellular responses. Upon TNFA stimulation RIPK1 is recruited to a TRADD-TRAF complex initiated by TNFR1 trimerization. There, it is ubiquitinated via 'Lys-63'-link chains, inducing its association with the IKK complex, and its activation through NEMO binding of polyubiquitin chains.

    • Sequence similarities

      Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
      Contains 1 death domain.
      Contains 1 protein kinase domain.

    • Post-translational
      modifications

      Proteolytically cleaved by caspase-8 during TNF-induced apoptosis. Cleavage abolishes NF-kappa-B activation and enhances pro-apototic signaling through the TRADD-FADD interaction.
      Autophosphorylated on serine and threonine residues.
      Ubiquitinated by 'Lys-11'-, 'Lys-48'-, 'Lys-63'- and linear-linked type ubiquitin. Polyubiquitination with 'Lys-63'-linked chains by TRAF2 induces association with the IKK complex. Deubiquitination of 'Lys-63'-linked chains and polyubiquitination with 'Lys-48'-linked chains by TNFAIP3 leads to RIPK1 proteasomal degradation and consequently to the termination of the TNF- or Linear polyubiquitinated; the head-to-tail polyubiquitination is mediated by the LUBAC complex. LPS-mediated activation of NF-kappa-B. Also ubiquitinated with 'Lys-11'-linked chains.

    • Cellular localization

      Cytoplasm.
    • Information by UniProt

    • Database links

      • Alternative names

        • Cell death protein RIP antibody
        • FLJ39204 antibody
        • OTTHUMP00000039163 antibody
        • Receptor (TNFRSF) interacting serine threonine kinase 1 antibody
        • receptor interacting protein 1 antibody
        • Receptor interacting protein antibody
        • Receptor interacting protein kinase 1 antibody
        • Receptor interacting serine threonine protein kinase 1 antibody
        • Receptor TNFRSF interacting serine threonine kinase 1 antibody
        • Receptor-interacting protein 1 antibody
        • Receptor-interacting serine/threonine-protein kinase 1 antibody
        • Rinp antibody
        • RIP 1 antibody
        • RIP antibody
        • Rip-1 antibody
        • RIP1 antibody
        • RIPK 1 antibody
        • Ripk1 antibody
        • RIPK1_HUMAN antibody
        • Serine threonine protein kinase RIP antibody
        • Serine/threonine-protein kinase RIP antibody
        see all

      Images

      • Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)
        Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)

        Lane 1: Wild-type HAP1 cell lysate (20 µg)
        Lane 2: RIP knockout HAP1 cell lysate (20 µg)
        Lane 3: HeLa cell lysate (20 µg)
        Lane 4: Raji cell lysate (20 µg)
        Lanes 1 to 4: Merged signal (red and green). Green - ab178420 observed at 78 kDa. Red - loading control, ab8245, observed at 37 kDa.


        ab178420 was shown to react with RIP in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when RIP knockout samples were examined. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab178420 and ab8245 (loading control to GAPDH) were both diluted 1/1000 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 hour at room temperature before imaging.

      • Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)
        Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)
        All lanes : Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution

        Lane 1 : Raji cell lysate
        Lane 2 : HeLa cell lysate with Staurosporine
        Lane 3 : HeLa cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP at 1/2000 dilution

        Predicted band size: 75 kDa

      • Flow Cytometry - Anti-RIP antibody [EPR4689-100] (ab178420)
        Flow Cytometry - Anti-RIP antibody [EPR4689-100] (ab178420)

        Flow Cytometric analysis of permeabilized HeLa cells labeling RIP with ab178420 at 1/10 dilution (red) compared with a rabbit IgG negative control (green).

      References

      Publishing research using ab178420? Please let us know so that we can cite the reference in this datasheet.

      ab178420 has not yet been referenced specifically in any publications.

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