Overview

  • Product name
    Anti-Ubiquitin antibody [EPR8589]
    See all Ubiquitin primary antibodies
  • Description
    Rabbit monoclonal [EPR8589] to Ubiquitin
  • Host species
    Rabbit
  • Specificity
    ab137031 recognizes polyubiquitin chains only.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WBmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in Human Ubiquitin (UniProt ID: P0CG48)

  • Positive control
    • HeLa or 293T cell lysates; Jurkat cells.
  • General notes

    A trial size is available to purchase for this antibody.

     

    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 ab137031 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
ICC/IF 1/250 - 1/500.
Flow Cyt 1/100 - 1/1000.

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

 

WB 1/1000 - 1/10000. Predicted molecular weight: 8 kDa. Molecular weight provided is for monoubiquitin
  • Application notes
    Is unsuitable for IHC-P or IP.
  • Target

    • Relevance
      Function: Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. Similarity: Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.
    • Cellular localization
      Cell Membrane, Cytoplasmic and Nuclear
    • Database links
    • Alternative names
      • Epididymis secretory protein Li 50 antibody
      • FLJ25987 antibody
      • HEL S 50 antibody
      • MGC8385 antibody
      • Polyubiquitin B antibody
      • RPS 27A antibody
      • RPS27A antibody
      • UBA 52 antibody
      • UBA 80 antibody
      • UBA52 antibody
      • UBA80 antibody
      • UBB antibody
      • UBB_HUMAN antibody
      • UBC antibody
      • UBCEP 1 antibody
      • UBCEP 2 antibody
      • UBCEP1 antibody
      • UBCEP2 antibody
      • Ubiquitin antibody
      • Ubiquitin B antibody
      see all

    Images

    • All lanes : Anti-Ubiquitin antibody [EPR8589] (ab137031) at 1/200 dilution

      Lane 1 : MCF-7 Whole Cell Lysate
      Lane 2 : MCF-7 Whole Cell Lysate + M132 (50 uM 90 min)
      Lane 3 : Mouse Brain

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 8 kDa



      This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. ab137031 and ab8245 (loading control to GAPDH) were diluted 1/200  and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging using the Licor Odyssey CLx.

    • Overlay histogram showing HepG2 cells stained with ab137031 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab137031, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • All lanes : Anti-Ubiquitin antibody [EPR8589] (ab137031) at 1/1000 dilution

      Lane 1 : Hela cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP conjugated Goat anti Rabbit IgG at 1/2000 dilution

      Predicted band size: 8 kDa

    • Immunofluorescence analysis of Jurkat cells labelling Ubiquitin with ab137031 at 1/250 dilution.
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    ab137031 has not yet been referenced specifically in any publications.

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