• Product nameAnti-Ubiquitin antibody [Ubi-1]
    See all Ubiquitin primary antibodies
  • Description
    Mouse monoclonal [Ubi-1] to Ubiquitin
  • Tested applicationsSuitable for: IHC-P, WB, ELISA, ICC/IF, IHC-Frmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Drosophila C virus
  • Immunogen

    Full length protein corresponding to Ubiquitin conjugated to Keyhole Limpet Haemocyanin (KLH). Mouse monoclonal [Ubi-1] to Ubiquitin was raised against purified ubiquitin conjugated with glutaraldehyde to keyhole limpet hemocyanin.

  • Positive control
    • H9C2 cells IHC-P:FFPE mouse brain alzheimer
  • General notes

    This product was changed from ascites to tissue culture supernatant on 17th May 2016. The following lots are from ascites and are still in stock as of 17th May 2016: GR159581-2, GR159581-12, GR159581-13. Lot numbers higher than GR159581-13 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.



Our Abpromise guarantee covers the use of ab7254 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
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/100 - 1/5000.
ELISA Use at an assay dependent concentration.
ICC/IF 1/500.
IHC-Fr 1/1000.
  • Application notesIs unsuitable for IP.
  • Target

    • 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
    • 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

    Anti-Ubiquitin antibody [Ubi-1] images

    • All lanes : Anti-Ubiquitin antibody [Ubi-1] (ab7254) at 1/100 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : HeLa cell lysate with MG132
      Lane 3 : Mouse brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Anti mouse IgG at 1/10000 dilution
    • IHC image of ab7254 staining in mouse alzheimer brain formalin fixed paraffin embedded tissue section, using MOM detection kit, ab127055. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab7254, 5µg/ml, for 15 mins at room temperature.  DAB was used as the chromogen (ab103723). The section was counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Immunohistochemical staining of ubiquitin in a 7 months-old MPS IIIA mouse brain (PFA fixed) using ab7254.

      Sections, 40 µm thick, were cut sagittally on a vibratome. Slices were permeabilized in 1% triton X-100/PBS for 1 hour at room temperature. The samples were placed on vectabond-coated glass slides and dried overnight. They were then rehydrated in PBS and washed in ice-cold methanol for 10 min.

      The slices were washed in 0.3% H2O2 in methanol at room temperature and incubated sequentially with 0.5% normal donkey serum, ab7254 and biotin-labeled F(ab')2 donkey secondary antibody in Tris-buffered saline (pH 7.5) containing 2% BSA and 0.02% Tween 20. The signal was detected with an ABC kit, visualized with diaminobenzidine and counterstained with hematoxylin.

    • ab7254 staining Ubiquitin in murine myocytes by Immunocytochemistry/ Immunofluorescence. Isolated myocytes were treated with 10µM FCCP for 1 hour, or untreated as the control. Cells were then fixed in formaldehyde, permeabilized using 1% Triton X-100, then blocked with 10% horse serum for 1 hour at room temperature, incubated with ab7254 at a 1/200 dilution. The secondary used was an Alexa-Fluor 488 conjugated goat anti-mouse polyclonal (ab150113), used at a 1/500 dilution.Results show that with FCCP treatment, Ubiquitin is recruited onto mitochondria.

      See Abreview

    • All lanes : Anti-Ubiquitin antibody [Ubi-1] (ab7254) at 1/500 dilution

      Lane 1 : Rat H9C2 whole cell lysated - untreated
      Lane 2 : Rat H9C2 whole cell lysated - treated with FCCP

      Lysates/proteins at 30 µg per lane.

      HRP-conjugated Goat anti-mouse IgG polyclonal at 1/4000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Exposure time : 2 minutes

      This image is courtesy of an anonymous Abreview

      See Abreview

    • Human normal placenta (ab29745). Staining is localised in the cytoplasm and in the nuclei. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus, at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

    • ab7254 staining Ubiquitin in Rat h9c2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 1% Triton X-100 and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/200 in 5% Goat serum ab7481) for 1 hour at room temperature. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (ab150113) (1/400) was used as the secondary antibody.

      See Abreview

    • Image courtesy of an anonymous Abreview.

      HeLa cells were co-transfected with a plasmid expressing a target protein together with Ubi expressing vector for 24 hours and either left untreated (Contr) or were treated with 10 µM MG-132 for 6 hours (+MG132). Then the protein of interest was pulled down using Flag agarose beads and and probed with ab7254 at a 1/2000 dilution. The secondary used was an Alexa-Fluor 680 conjugated goat anti-mouse polyclonal used at a 1/10000 dilution. The protein is known to be degraded through proteasome.

      See Abreview

    References for Anti-Ubiquitin antibody [Ubi-1] (ab7254)

    This product has been referenced in:
    • Yu Z  et al. Lys29-linkage of ASK1 by Skp1-Cullin 1-Fbxo21 ubiquitin ligase complex is required for antiviral innate response. Elife 5:N/A (2016). Read more (PubMed: 27063938) »
    • Li Q  et al. Temporal Profiles of Stress Protein Inductions after Focal Transient Ischemia in Mice Brain. J Stroke Cerebrovasc Dis 25:2344-51 (2016). Read more (PubMed: 27318648) »

    See all 21 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (Cardiomyocytes)
    Gel Running Conditions Reduced Denaturing
    Loading amount 25 µg
    Treatment Starvation
    Specification Cardiomyocytes
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Nov 11 2016

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Sample Human Cell (Human primary fibroblasts)
    Specification Human primary fibroblasts
    Permeabilization Yes - 0.2% Triton-X100
    Fixative Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Mar 26 2014

    Thank you for your enquiry.

    The immunogen was purified bovine ubiquitin isolated from erythrocytes and cross-linked into an insoluble complex by glutaraldehyde. Ubiquitin is one of the most conserved proteins and the sequence is virtually id...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Drosophila C Virus Cell (fly heart tube)
    Specification fly heart tube
    Fixative Paraformaldehyde
    Permeabilization Yes - 1% triton-x100
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Feb 11 2013

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

    Read More

    Thank you for taking the time to complete the questionnaire and provide the order number. I am sorry to hear the customer has had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will provide...

    Read More

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab7254.

    To check the status of the order please contact ou...

    Read More

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab7254.

    To check the status of the order please contact our...

    Read More

    Thanks for your email. ab4587 does not have any required MSDS information, so a link will not be present on this datasheet. For example, ab7254 does have a link to MSDS information. I hope this helps. Please contact me again if you have any further que...

    Read More

    Thanks for your enquiry. If required, MSDS information is located via one or more 'MSDS' links near the top left corner of the online datasheet. This link is just below the 'Datasheet PDF' link present on all datasheets. I hope this is helpful. Please ...

    Read More

    1-10 of 28 Abreviews or Q&A