Overview

  • Product name
  • Description
    Rabbit polyclonal to Ubiquitin
  • Specificity
    It can identify free ubiquitin as well as ubiquinated proteins. The antibody recognizes polyubiquitin chains more strongly than monoubiquitinated molecules.
  • Tested applications
    Suitable for: Immunoelectrophoresis, IHC-FoFr, ICC/IF, WB, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Saccharomyces cerevisiae, Xenopus laevis, Drosophila melanogaster, Fish, Monkey, Escherichia coli
  • Immunogen

    Full length native protein (purified) corresponding to Cow Ubiquitin conjugated to Keyhole Limpet Haemocyanin (KLH).

  • Positive control
    • Heat-shocked HeLa cell lysate, mouse brain tissue extract.

Properties

Applications

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.

Application Abreviews Notes
Immunoelectrophoresis 1/1000.
IHC-FoFr 1/200.
ICC/IF Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Predicted molecular weight: 10 kDa.
IP 1/100. PubMed: 20103532
  • Application notes
    Is unsuitable for IHC-P.
  • 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

    • Lane 1 :
      Lane 2 :
      Lane 3 :

      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.

    References

    This product has been referenced in:
    • Luo X  et al. Caveolin 1-related autophagy initiated by aldosterone-induced oxidation promotes liver sinusoidal endothelial cells defenestration. Redox Biol 13:508-521 (2017). Read more (PubMed: 28734243) »
    • Yang J  et al. Spatial sequestration and detoxification of Huntingtin by the ribosome quality control complex. Elife 5:N/A (2016). Read more (PubMed: 27033550) »

    See all 19 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (spermatocytes)
    Permeabilization
    No
    Specification
    spermatocytes
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 27°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Dec 16 2016

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (EC CELLS)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    60 µg
    Specification
    EC CELLS
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Username

    Abcam user community

    Verified customer

    Submitted Apr 08 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse mammary cell)
    Loading amount
    250 µg
    Specification
    Mouse mammary cell
    Treatment
    5ng/ml TGF beta for 8hrs
    Gel Running Conditions
    Reduced Denaturing (10.0% gel)
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 07 2011

    Thank you for your enquiry. Further to correspondence with the source of this antibody I can tell you that the following approach was employed for the testing of this antibody by western blot analysis. In brief the samples were prepared in denaturi...

    Read More
    Application
    Western blot
    Sample
    African Green Monkey Cell lysate - whole cell (Vero)
    Loading amount
    30000 cells
    Specification
    Vero
    Treatment
    CO-IP for the prot of interest (40kDa)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10
    Username

    Mr. olivier Reynard

    Verified customer

    Submitted Jul 27 2006

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunoprecipitation
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (Saccharomyces cerevisiae)
    Total protein in input
    5 µg
    Specification
    Saccharomyces cerevisiae
    Immuno-precipitation step
    Protein G
    Username

    Abcam user community

    Verified customer

    Submitted Jun 19 2006

    Thank you for your enquiry. It has not been determined whether ab14372 detects mono- or poly-ubiquitinated proteins. It would quite likely recognize both. Ab19247 should recognize ubiquinated proteins in Western blots regardless of how may ubiq...

    Read More

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