Recombinant
RabMAb

Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)

Overview

  • Product name
    Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448]
    See all Ubiquitin primary antibodies
  • Description
    Rabbit monoclonal [EPR8590-448] to Ubiquitin (linkage-specific K63)
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IHC-FoFr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Ubiquitin aa 50-150 (linkage-specific K63) (Cysteine residue). The exact sequence is proprietary.
    Database link: P0CG48

  • Positive control
    • WB: K63-linked-Ub2-7 recombinant protein. HEk-293 and HeLa cell lysate. Mouse and rat brain lysate. IHC: Human kidney carcinoma tissue. Human tonsil tissue. FC: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels.

    If you have any questions regarding this update, please contact our Scientific Support team.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Applications

Our Abpromise guarantee covers the use of ab179434 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
WB 1/1000. Predicted molecular weight: 77 kDa.
IHC-P 1/250 - 1/500.

For unpurified, use 1/100 - 1/250.

ICC/IF 1/430.

For unpurified, use 1/100 - 1/250.

IHC-FoFr Use at an assay dependent concentration. PubMed: 28243725
Flow Cyt 1/210.

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

  • All lanes : Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434) at 1/1000 dilution (purified)

    Lane 1 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 77 kDa
    Observed band size : 16-300 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human tonsil with purified ab179434 at a dilution of 1/500. The secondary antibody used is a HRP conjugated goat anti-rabbit (ab97051, 1/500). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • All lanes : Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434) at 1/5000 dilution (purified)

    Lane 1 : K6-linked-Ub2 recombinant protein
    Lane 2 : K11-linked-Ub2 recombinant protein
    Lane 3 : K29-linked-Ub2 recombinant protein
    Lane 4 : K33-linked-Ub2 recombinant protein
    Lane 5 : K48-linked-Ub2 recombinant protein
    Lane 6 : K63-linked-Ub2 recombinant protein

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 77 kDa
    Observed band size : 16-70 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Ubiquitin (linkage-specific K63) with purified ab179434 at 1/210 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • All lanes : Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434) at 1/1000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 77 kDa
    Observed band size : 16-300 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded human kidney carcinoma tissue labeling K63-linkage specific polyubiquitin with unpurified ab179434 at 1/100 dilution.

  • All lanes : Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434) at 1/1000 dilution (unpurified)

    Lane 1 : Ubiquitin recombinant protein
    Lane 2 : K6-linked-Ub2 recombinant protein
    Lane 3 : K11-linked-Ub2 recombinant protein
    Lane 4 : K27-linked-Ub2 recombinant protein
    Lane 5 : K29-linked-Ub2 recombinant protein
    Lane 6 : K33-linked-Ub2 recombinant protein
    Lane 7 : K48-linked-Ub2-7 recombinant protein
    Lane 8 : K63-linked-Ub2-7 recombinant protein

    Lysates/proteins at 0.02 µg per lane.


    Predicted band size : 77 kDa

References

This product has been referenced in:
  • Li X  et al. USP9X regulates centrosome duplication and promotes breast carcinogenesis. Nat Commun 8:14866 (2017). WB . Read more (PubMed: 28361952) »
  • Scekic-Zahirovic J  et al. Motor neuron intrinsic and extrinsic mechanisms contribute to the pathogenesis of FUS-associated amyotrophic lateral sclerosis. Acta Neuropathol 133:887-906 (2017). IHC-FoFr ; Mouse . Read more (PubMed: 28243725) »

See all 8 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 20°C
Sample
Mouse Cell (immortalised bone marrow-derived macrophages)
Specification
immortalised bone marrow-derived macrophages
Permeabilization
Yes - 0.1% Triton X-100
Fixative
Paraformaldehyde
Username

Dr. James Harris

Verified customer

Submitted Sep 08 2014

Application
Western blot
Loading amount
0.03 µg
Gel Running Conditions
Reduced Denaturing (4-12)
Sample
Human Recombinant protein (Recombinant protein)
Specification
Recombinant protein
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 19 2013

Application
Western blot
Loading amount
0.01 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Recombinant protein (recombinant protein)
Specification
recombinant protein
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Nov 12 2013

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

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