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  1. Link

    products/primary-antibodies/ubiquitin-linkage-specific-k63-antibody-epr8590-448-ab179434.pdf

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RecombinantRabMAb

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

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  • SDS
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Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Flow Cytometry (Intracellular) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
  • Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR8590-448] to Ubiquitin (linkage-specific K63)
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP PE

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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)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide 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 lysates, and Mouse and rat brain lysate. IHC-P: Human kidney carcinoma and tonsil tissue. Flow Cyt (intra): HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped 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 buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8590-448
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Tangles & Tau
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Ubiquitylation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • Other
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • Ubiquitin like Modifiers
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab192489)
    • Alexa Fluor® 488 Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab192539)
    • Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] - Low endotoxin, Azide free (ab220794)
    • Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] - BSA and Azide free (ab271929)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/210.
WB (4)
1/1000. Predicted molecular weight: 77 kDa.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

Notes
Flow Cyt (Intra)
1/210.
WB
1/1000. Predicted molecular weight: 77 kDa.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

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

    • Entrez Gene: 7314 Human
    • Entrez Gene: 22187 Mouse
    • Entrez Gene: 192255 Rat
    • SwissProt: P0CG47 Human
    • SwissProt: P0CG48 Human
    • SwissProt: P62979 Human
    • SwissProt: P62987 Human
    • SwissProt: P62988 Human
    • SwissProt: P0CG49 Mouse
    • SwissProt: P62991 Mouse
    • SwissProt: P0CG51 Rat
    • SwissProt: P62989 Rat
    • Unigene: 356190 Human
    • Unigene: 520348 Human
    • Unigene: 282093 Mouse
    • Unigene: 371592 Mouse
    • Unigene: 110618 Rat
    • Unigene: 1253 Rat
    • Unigene: 3761 Rat
    see all
  • 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

  • Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    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
    All lanes : 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)

    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.

  • Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    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
    All lanes : 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 (Intracellular) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    Flow Cytometry (Intracellular) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)

    Intracellular 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. 

     

     

  • Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    Western blot - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    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
    All lanes : 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (ab179434)

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

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

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

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (52)

Publishing research using ab179434? Please let us know so that we can cite the reference in this datasheet.

ab179434 has been referenced in 52 publications.

  • Tan CT  et al. MOAP-1-mediated dissociation of p62/SQSTM1 bodies releases Keap1 and suppresses Nrf2 signaling. EMBO Rep 22:e50854 (2021). PubMed: 33393215
  • Liu C  et al. Cytoplasmic SHMT2 drives the progression and metastasis of colorectal cancer by inhibiting ß-catenin degradation. Theranostics 11:2966-2986 (2021). PubMed: 33456583
  • Nakabayashi O  et al. MIND bomb 2 prevents RIPK1 kinase activity-dependent and -independent apoptosis through ubiquitylation of cFLIPL. Commun Biol 4:80 (2021). PubMed: 33469115
  • Zeng Y  et al. The PB1 protein of influenza A virus inhibits the innate immune response by targeting MAVS for NBR1-mediated selective autophagic degradation. PLoS Pathog 17:e1009300 (2021). PubMed: 33577621
  • Ranes M  et al. Reconstitution of the destruction complex defines roles of AXIN polymers and APC in ß-catenin capture, phosphorylation, and ubiquitylation. Mol Cell 81:3246-3261.e11 (2021). PubMed: 34352208
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Dopaminergic Neurons)
Gel Running Conditions
Reduced Denaturing (NuPage 4-12% MES buffer)
Loading amount
30 µg
Specification
Dopaminergic Neurons
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 20 2020

Western blot abreview for Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (colon cancer, kidney)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
colon cancer, kidney
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 09 2020

ELISA abreview for Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448]

Excellent
Abreviews
Abreviews
abreview image
Application
ELISA
Sample
Rat Serum (SD rat serum)
Specification
SD rat serum
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Type
Sandwich (Capture)
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 24 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. James Harris

Verified customer

Submitted Sep 08 2014

Western blot abreview for Anti-Ubiquitin (linkage-specific K63) [EPR8590-448] antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 19 2013

Western blot abreview for Anti-Ubiquitin (linkage-specific K63) [EPR8590-448] antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 12 2013

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