Overview

  • Product nameAnti-Ubiquitin antibody
    See all Ubiquitin primary antibodies
  • Description
    Rabbit polyclonal to Ubiquitin
  • SpecificityThis antibody reacts with ubiquitin, a polypeptide w/ Mr. of approx. 8.5kD. Reacts with physiologically occurring conjugates of ubiquitin and intracellular proteins. Specifically recognizes ubiquitinated cytoplasmic inclusion bodies.
  • Tested applicationsICC/IF, IHC-FoFr, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African Green Monkey
    Predicted to work with: Horse, Cow
  • Immunogen

    Recombinant full length protein (Human)

  • Positive control
    • Pancreas and Alzheimer’s brain Rat brain
  • General notes

    This antibody stains the periphery of senile plaques and of neurofibrillary tangles in Alzheimer's disease brain, the Lewy bodies in Parkinson's disease brain, and the Mallory bodies in alcoholic liver disease.

Properties

Applications

Our Abpromise guarantee covers the use of ab7780 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 Use at an assay dependent concentration.

1/500 - 1/1000.

IHC-FoFr 1/50 - 1/100.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Boil tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.
WB Use at an assay dependent concentration. Predicted molecular weight: 8.5 kDa. Used at a dilution of 1/2000 B16F10 murine melanoma cell line (see Abreview). Predicted molecular weight: 8.5 kDa.
IP Use at an assay dependent concentration.

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
    • FLJ25987 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 images


  • developed using the ECL technique

    Predicted band size : 8.5 kDa


    Exposure time : 30 seconds

    Image courtesy of an anonymous Abreview.

    ab7780 used in Western Blot. Whole cell lysate prepared from human lung cancer cell lines was loaded at 100000 cells. The gel was run under reduced, denaturing conditions. 5% milk used as the blocking agent. ab7780 used at a 1/1000 dilution, incubated for 16 hours at 4°C in 5% BSA in TBS-Tween (0.1%). The secondary was an HRP conjugated mouse anti-rabbit, used at a 1/10,000 dilution.Specific bands seen : 16, 24, 32, 50, 60, kDa.

  • All lanes : Anti-Ubiquitin antibody (ab7780) at 1/2000 dilution

    Lane 1 : Human Panc-1 whole cell lysate
    Lane 2 : Human Panc-1 whole cell lysate

    Lysates/proteins at 700 µg per lane.

    Secondary
    HRP Donkey anti-rabbit IgG polyclonal at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 8.5 kDa


    Exposure time : 25 seconds

    This image is courtesy of an Abreview submitted by Armen Petrosyan

    Ubiquitin western blot of the complexes pulled down with anti-c-Myc antibodies from the lysates of cells treated with DMSO or MG-132. Treatment with the proteasome inhibitor, MG-132, increases ubiquitinated protein; seen as several bands between 55-72 kDa.

    See Abreview

  • ab7780 staining Ubiquitin in Panc-1 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in formaldehyde, blocked with 1% serum for 1 hour at 22°C and then incubated with ab7780 at a 1/2000 dilution for 1 hour at 22°C. The secondary was used at a 1/200 dilution.Green - Golgi residental proteins C2GnT-M.Red -Ubiquitin.Blue - nucleus staining by DAPI.

    See Abreview

  • ICC/IF image of ab7780 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7780, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) ab150113) used at a 1/1000 dilution for 1h. Alexa Fluor© 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43æM.

  • ab7780 at a dilution of 1/1000, staining Ubiquitin in neurons (Alexa 488 secondary at 1/2000) on 30µm coronal rat brain (ab29475) tissue sections in free floating IHC (see protocol link for detailed description). Image shows neuronal staining observed with [A] 20x objective and [B] 40x objective.

    NB: No labeling observed following omission of primary antibody.

    Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

  • ab7780 (1/100) staining Ubiquitin in paraffin-embedded brain sections of a patient with Alzheimer's disease.

References for Anti-Ubiquitin antibody (ab7780)

This product has been referenced in:
  • Shrivastava V  et al. Cigarette smoke affects posttranslational modifications and inhibits capacitation-induced changes in human sperm proteins. Reprod Toxicol 43:125-9 (2014). Read more (PubMed: 24345728) »
  • Xu YH  et al. Multiple pathogenic proteins implicated in neuronopathic Gaucher disease mice. Hum Mol Genet N/A:N/A (2014). Mouse . Read more (PubMed: 24599400) »

See all 14 Publications for this product

Product Wall

Application Western blot
Loading amount 1000 µg
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (Brain cancer cells (immunoprecipitated lysates))
Specification Brain cancer cells (immunoprecipitated lysates)
Treatment proteasome inhibitor MG-132
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Human Cell (HS 5)
Specification HS 5
Permeabilization Yes - Triton X 100
Fixative Formaldehyde
Username

Mr. Kannan Govindaraj

Verified customer

Submitted Aug 02 2013



We do not have any experimental data regarding the cross-reactivity of ab7780 with SUMO proteins. I compared the immunogen for ab7780 with the sequences of SUMO 1, 2, 3, 4 (P63165, P61956, P55854, Q6EEV6-Uniprot) and the percentage homology i...

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I am happy to hear that the antibody is working for you when using BSA instead of milk! If you have any additional questions, please let me know and I will be happy to help you further.



Any products that require an MSDS form will have a link on the product's online datasheet to the data. It will be a PDF document link labeled 'SDS'. Please see attached image for example of what to look for on the datasheet. If no 'SDS' link ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Skeletal muscle (AT))
Loading amount 30 µg
Specification Skeletal muscle (AT)
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. G.K Sakellariou

Verified customer

Submitted Sep 28 2012

Thank you for your reply.
This antibody should bind to both the 8.5 kDa free ubiquitin and any other ubiquitinated proteins in your sample. For that reason, you may observe multiple bands in WB. If you are looking to see if one protein in particul...

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Thank you for contacting us. I am sorry that the replacement antibody is also giving no signal in WB. Ubiquitin should be expressed in all tissues and cell types, but it is possible that the protein is not being effectively released from your samples. ...

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Thank you for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial of ab7254 on the order ***, which should arrive tomorrow.

Please keep me updated about th...

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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 ab20137 and one vial of ab131107.

Expect this shipment to a...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"