Recombinant Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab181537)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17034] to Ubiquitin (linkage-specific K27)
- Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034]
See all Ubiquitin primary antibodies -
Description
Rabbit monoclonal [EPR17034] to Ubiquitin (linkage-specific K27) -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: K27-linked-Ub2 recombinant protein. C6, RAW 264.7, PC-12, NIH/3T3, HeLa, 293T and Jurkat whole cell lysates. Rat and Mouse brain, heart, kidney and spleen lysates. IHC-P: Human transitional cell carcinoma of bladder tissue. Mouse and Rat spleen tissue. Flow Cyt (intra): Jurkat 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.
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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17034 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab237254)
- Alexa Fluor® 647 Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab237255)
- HRP Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab237256)
- Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] - BSA and Azide free (ab238442)
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181537 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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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/5000.
Actual Band: 14kDa and above. |
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Flow Cyt (Intra) |
1/160.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/500.
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Notes |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/5000. Actual Band: 14kDa and above. |
Flow Cyt (Intra)
1/160. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/500. |
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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Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma)
labelling Ubiquitin (linkage-specific K27) with purified ab181537 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control. -
All lanes : Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab181537) at 1/5000 dilution
Lane 1 : K27-linked-Ub2 recombinant protein
Lane 2 : K6-linked-Ub2 recombinant protein
Lane 3 : K11-linked-Ub2 recombinant protein
Lane 4 : K29-linked-Ub2 recombinant protein
Lane 5 : K33-linked-Ub2 recombinant protein
Lane 6 : K48-linked-Ub2 recombinant protein
Lane 7 : K63-linked-Ub2 recombinant protein
Lane 8 : Ubiquitin recombinant protein
Lane 9 : Ubiquitin linear recombinant protein
Lysates/proteins at 0.02 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilutionBlocking/dilution buffer: 5% BSA/TBST.
Expected MW is ~17kDa (8.5*2)
The ~30kDa band may be the dimer of K27-linked-Ub2 recombinant protein.
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All lanes : Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab181537) at 1/50000 dilution
Lane 1 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Observed band size: 26 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% BSA/TBST.
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Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab181537) at 1/5000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Observed band size: 26 kDa why is the actual band size different from the predicted?Blocking/dilution buffer: 5% BSA/TBST.
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All lanes : Anti-Ubiquitin (linkage-specific K27) antibody [EPR17034] (ab181537) at 1/5000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Mouse heart lysates
Lane 3 : Mouse kidney lysates
Lane 4 : Mouse spleen lysates
Lane 5 : Rat brain lysates
Lane 6 : Rat heart lysates
Lane 7 : Rat kidney lysates
Lane 8 : Rat spleen lysates
Lane 9 : C6 (Rat glial tumor cells) whole cell lysates
Lane 10 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 12 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Observed band size: 14,26 kDa why is the actual band size different from the predicted?Blocking/dilution buffer: 5% BSA/TBST.
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Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder labeling Ubiquitin (linkage-specific K27) with ab181537 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on cancer cells of Human bladder transitional cell carcinoma is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Ubiquitin (linkage-specific K27) with ab181537 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on lymphocytes of Mouse spleen is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Ubiquitin (linkage-specific K27) with ab181537 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on lymphocytes of Rat spleen is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of2% paraformaldehyde-fixedJurkat (Human T cell leukemia cells from peripheral blood) cells labeling Ubiquitin (linkage-specific K27) with ab181537 at 1/160 dilution (red). The secondary antibody wasGoat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control isRabbit monoclonal IgG (black) and cell without incubation with primary antibody and secondary antibody is blue.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (14)
ab181537 has been referenced in 14 publications.
- Zittlau KI et al. Temporal Analysis of Protein Ubiquitylation and Phosphorylation During Parkin-Dependent Mitophagy. Mol Cell Proteomics 21:100191 (2022). PubMed: 34974192
- Shearer RF et al. K27-linked ubiquitylation promotes p97 substrate processing and is essential for cell proliferation. EMBO J 41:e110145 (2022). PubMed: 35349166
- Li F et al. Targeting HECTD3-IKKα axis inhibits inflammation-related metastasis. Signal Transduct Target Ther 7:264 (2022). PubMed: 35918322
- Zhang X et al. Extracellular fibrinogen-binding protein released by intracellular Staphylococcus aureus suppresses host immunity by targeting TRAF3. Nat Commun 13:5493 (2022). PubMed: 36123338
- Chen X et al. OTUD1 Regulates Antifungal Innate Immunity through Deubiquitination of CARD9. J Immunol 206:1832-1843 (2021). PubMed: 33789983