Overview

  • Product name

    Anti-RIP antibody [EPR19697]
    See all RIP primary antibodies
  • Description

    Rabbit monoclonal [EPR19697] to RIP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse RIP aa 300-600. The exact sequence is proprietary.
    Database link: Q60855

  • Positive control

    • WB: NIH/3T3 and RAW 264.7 whole cell lysates; mouse brain, heart and kidney lysates. ICC/IF: NIH/3T3 and RAW 264.7 cells. Flow Cyt: NIH/3T3 and RAW 264.7 cells. IP: NIH/3T3 whole cell lysate.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab202985 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. Detects a band of approximately 75, 44 kDa (predicted molecular weight: 75 kDa).
ICC/IF 1/200.
Flow Cyt 1/500.
IP 1/30.

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

  • All lanes : Anti-RIP antibody [EPR19697] (ab202985) at 1/1000 dilution

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Mouse kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 75 kDa
    Observed band size: 44,75 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 8 seconds; Lane 2: 15 seconds; Lane 3-5: 3 minutes.

    The molecular weight observed is consistent with what has been described in the literatures  (PMID: 15314284 & 10521396).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling RIP with ab202985 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The results show cytoplasmic staining on NIH/3T3 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab202985 at 1/200 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling RIP with ab202985 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The results show cytoplasmic staining on RAW 264.7 cells. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab202985 at 1/200 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling RIP with ab202985 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling RIP with ab202985 at 1/500 dilution (red) compared with a Rabbit IgG,monoclonal[EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • ab202985 Immunoprecipitating RIP in mouse embryo whole cell lysate. 2µg of capture antibody per 0.35mg of lysate was used. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/1000 and VeriBlot for IP Detection Reagent (HRP) ab131366 was used for detection of 1/1000.

    Lane 1: NIH/3T3 (mouse embryo) whole cell lysate
    Lane 2: NIH/3T3 (mouse embryo) whole cell lysate 
    Lane 3: Rabbit monoclonal IgG ab172730 instead of ab202985 in NIH/3T3 (mouse embryo) whole cell lysate

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

References

This product has been referenced in:

  • Zhang ZX  et al. Intracellular pH Regulates TRAIL-Induced Apoptosis and Necroptosis in Endothelial Cells. J Immunol Res 2017:1503960 (2017). WB ; Mouse . Read more (PubMed: 28884134) »
See 1 Publication for this product

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