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  1. Link

    rip-antibody-epr4689-100-ab178420.pdf

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Cell Biology Apoptosis Intracellular Kinases
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
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Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)
  • Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)
  • Flow Cytometry (Intracellular) - Anti-RIP antibody [EPR4689-100] (ab178420)
  • Anti-RIP antibody [EPR4689-100] (ab178420)

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: Flow Cyt (Intra), WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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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: Flow Cyt (Intra), WBmore 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

    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.

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

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4689-100
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Kinases
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Death receptors & ligands
    • RIP
    • Cancer
    • Cell Death
    • Apoptosis
    • Receptors
    • Death receptors & ligands
    • RIP
    • Cancer
    • Cell Death
    • Necroptosis
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-RIP antibody [EPR4689-100] - BSA and Azide free (ab227843)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Raji whole cell lysate (ab30124)
  • Recombinant Protein

    • Recombinant human RIP protein (Active) (ab190411)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/10 - 1/100.

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

WB
1/1000 - 1/10000. Predicted molecular weight: 75 kDa.
Notes
Flow Cyt (Intra)
1/10 - 1/100.

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

WB
1/1000 - 1/10000. Predicted molecular weight: 75 kDa.
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.
  • Target information above from: UniProt accession Q13546 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8737 Human
    • Omim: 603453 Human
    • SwissProt: Q13546 Human
    • Unigene: 519842 Human
    • 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 (Intracellular) - Anti-RIP antibody [EPR4689-100] (ab178420)
      Flow Cytometry (Intracellular) - Anti-RIP antibody [EPR4689-100] (ab178420)

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

       

       

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

    Protocols

    • Western blot protocols
    • Flow cytometry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (1)

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

    ab178420 has been referenced in 1 publication.

    • Moriwaki K  et al. The death-inducing activity of RIPK1 is regulated by the pH environment. Sci Signal 13:N/A (2020). PubMed: 32398349

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