Recombinant Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (ab247278)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP296Y] to Ubiquitin - BSA and Azide free
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free
See all Ubiquitin primary antibodies -
Description
Rabbit monoclonal [EP296Y] to Ubiquitin - BSA and Azide free -
Host species
Rabbit -
Specificity
The antibody recognizes only mono-ubiquitinated molecules.
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Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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General notes
ab247278 is the carrier-free version of ab33893.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP296Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247278 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 |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 8.5 kDa (predicted molecular weight: 8 kDa).
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Detects a band of approximately 8.5 kDa (predicted molecular weight: 8 kDa). |
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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (ab247278)This data was developed using ab33893, the same antibody clone in a different buffer formulation.IHC image of Ubiquitin staining in mouse Alzheimer brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab33893, 1in500, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-Ubiquitin antibody [EP296Y] (ab33893) at 1/1000 dilution
Lane 1 : MCF-7 Whole cell lysate
Lane 2 : MCF-7 Whole cell lysate + (50 uM 90 min)
Lane 3 : Mouse brain tissue lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 8 kDaThis data was developed using ab33893, the same antibody clone in a different buffer formulation.
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. ab33893 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging using the Licor Odyssey CLx.
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Anti-Ubiquitin antibody [EP296Y] (ab33893) at 1/2000 dilution + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Predicted band size: 8 kDa
Observed band size: 8.5 kDa why is the actual band size different from the predicted?This data was developed using ab33893, the same antibody clone in a different buffer formulation.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (ab247278)This data was developed using ab33893, the same antibody clone in a different buffer formulation.ab33893 at a 1:250 dilution staining Ubiquitin in human prostate adenocarcinoma using Immunohistochemistry, Paraffin Embedded Tissue. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab247278 has not yet been referenced specifically in any publications.