Product nameAnti-Ubiquitin antibody
See all Ubiquitin primary antibodies
DescriptionRabbit polyclonal to Ubiquitin
SpecificityIt can identify free ubiquitin as well as ubiquinated proteins. The antibody recognizes polyubiquitin chains more strongly than monoubiquitinated molecules.
Tested applicationsSuitable for: Immunoelectrophoresis, IHC-FoFr, ICC/IF, WB, IPmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Mouse, Rat, Sheep, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Saccharomyces cerevisiae, Xenopus laevis, Drosophila melanogaster, Fish, Monkey, Escherichia coli
Full length native protein (purified) corresponding to Cow Ubiquitin conjugated to Keyhole Limpet Haemocyanin (KLH).
- Heat-shocked HeLa cell lysate, mouse brain tissue extract.
Storage instructionsShipped 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 bufferpH: 7.20
Preservative: 0.09% Sodium azide
Constituents: PBS, 50% Glycerol
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab19247 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/1000 - 1/5000. Predicted molecular weight: 10 kDa.|
|IP||1/100. PubMed: 20103532|
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 :
Lane 1 : HeLa cell lysate (Control, untreated)
Lane 2 : HeLa cells treated with MG132 (50 uM for 90 min)
Lane 3 : Mouse Brain
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 10 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. ab19247 and ab8245 (loading control to GAPDH) were diluted 1/200 and 1/10000 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.
Anti-Ubiquitin antibody (ab19247) at 1/1000 dilution + Heat-shocked HeLa cell lysate
Predicted band size: 10 kDa
Scrambled and lentivirus-stable infected SN4741 cells grown on coverslips were induced to differentiate under restrictive conditions for 24 h (pre-lethal stage), fixed and permeabilized with 0.1% Triton X-100 + 0.1% sodium citrate for 5 minutes. Cells were blocked with 10% donkey serum in PBS for 1 hour at 37°C and incubated over night at 4°C with rabbit polyclonal antibody against ubiquitin (ab19247, 1:100) and mouse monoclonal α-synuclein antibody (1:25). Samples were washed twice with PBS for 10 minutes before the addition of secondary antibodies at room temperature in the dark for 1 hour
Scrambled infected (control) cells show a diffuse staining pattern and do not show any inclusions at both permissive and restrictive temperatures (upper images). SKP1A-deficient cells developed multiple and perinuclear (bottom images, see arrows and insets) inclusion bodies 24 h after induction of differentiation, with characteristics of aggresomes. The reactivity of the aggregates to the different antibodies demonstrated a similar pattern as indicated by co-localized yellow immunostaining in the overlaid right-hand panel. No fluorescence was detected when the primary antibody was omitted. Each panel shows a representative picture of 10–15 views in two independent experiments.
Images top - bottom:
Scrambled 33°C/10% FCS,
SKP1A shRNA-1 33°C/10% FCS,
Scrambled 39°C/0.5% FCS,
SKP1A shRNA-1 39°C/0.5% FCS.
This product has been referenced in:
- Deng L et al. Ubiquitination of Rheb governs growth factor-induced mTORC1 activation. Cell Res 29:136-150 (2019). Read more (PubMed: 30514904) »
- Childers CL & Storey KB Purification and characterization of a urea sensitive lactate dehydrogenase from skeletal muscle of the African clawed frog, Xenopus laevis. J Comp Physiol B 189:271-281 (2019). Read more (PubMed: 30631901) »