Recombinant Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] - Low endotoxin, Azide free (ab221211)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP8589] to Ubiquitin (linkage-specific K48) - Low endotoxin, Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Overview
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Product name
Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] - Low endotoxin, Azide free
See all Ubiquitin primary antibodies -
Description
Rabbit monoclonal [EP8589] to Ubiquitin (linkage-specific K48) - Low endotoxin, Azide free -
Host species
Rabbit -
Specificity
This antibody only recognizes polyubiquitin chains formed by Lys-48 (K48) residue linkage. This antibody can detect the target in mouse and rat cell lines and induced tissues.
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore 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
- K48-linked-Ub2-7 This antibody gave a positive result when used in the following methanol fixed cell lines: MCF-7
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General notes
ab221211 is the carrier-free version of ab140601.
The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.
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 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.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
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
EP8589 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)
- Alexa Fluor® 488 Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab204274)
- Alexa Fluor® 647 Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab204476)
- Alexa Fluor® 594 Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab206366)
- Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] - BSA and Azide free (ab271911)
- PE Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab303017)
- APC Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab303018)
- HRP Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab303019)
- Alexa Fluor® 555 Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab312065)
- Alexa Fluor® 568 Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab312542)
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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 ab221211 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) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. |
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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Purified ab140601 staining Ubiquitin (linkage-specific K48) in MCF7 (Human breast adenocarcinoma cell line) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a counterstain for primary antibody ab133645 at 1/2000. DAPI was used as a nuclear counterstain and PBS as a negative control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140601).
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Purified ab140601 staining Ubiquitin (linkage-specific K48) in human endometrium carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140601).
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ICC/IF image of ab140601 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab140601 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140601).
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Overlay histogram showing HeLa cells stained with ab140601 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab140601, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140601).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab221211 has not yet been referenced specifically in any publications.