Product nameAnti-Ubiquitin antibody [EPR8589]
See all Ubiquitin primary antibodies
DescriptionRabbit monoclonal [EPR8589] to Ubiquitin
Specificityab137031 recognizes polyubiquitin chains only.
Tested applicationsSuitable for: ICC/IF, Flow Cyt, WBmore details
Unsuitable for: IHC-P or IP
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide corresponding to residues in Human Ubiquitin (UniProt ID: P0CG48)
- HeLa or 293T cell lysates; Jurkat cells.
A trial size is available to purchase for this antibody.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Dissociation constant (KD)KD = 4.95 x 10 -11 M Learn more about KD
Storage bufferPreservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
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.
|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|
RelevanceFunction: 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 localizationCell Membrane, Cytoplasmic and Nuclear
- Epididymis secretory protein Li 50 antibody
- FLJ25987 antibody
- HEL S 50 antibody
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.
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
ab137031 has not yet been referenced specifically in any publications.