Recombinant 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
Related conjugates and formulations
Overview
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR8590-448 -
Isotype
IgG -
Research areas
Associated products
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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)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
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 |
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Flow Cyt (Intra) |
1/210.
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WB | (4) |
1/1000. Predicted molecular weight: 77 kDa.
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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 |
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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
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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
see all -
Alternative names
- Epididymis secretory protein Li 50 antibody
- FLJ25987 antibody
- HEL S 50 antibody
see all
Images
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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
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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.
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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
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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.
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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
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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.
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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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (52)
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