Recombinant Anti-UBE2I / UBC9 antibody [EP2938Y] (ab75854)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2938Y] to UBE2I / UBC9
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-UBE2I / UBC9 antibody [EP2938Y]
See all UBE2I / UBC9 primary antibodies -
Description
Rabbit monoclonal [EP2938Y] to UBE2I / UBC9 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide within Human UBE2I/ UBC9 aa 1-100 (N terminal). The exact sequence is proprietary.
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Positive control
- U937, HeLa, Jurkat or HUVEC cell lysates. Human brain tissue. ICC/IF: 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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2938Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75854 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) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (4) |
1/1000 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
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IP | (1) |
Use at an assay dependent concentration.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100.
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Notes |
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Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa). |
IP
Use at an assay dependent concentration. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100. |
Target
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Function
Accepts the ubiquitin-like proteins SUMO1, SUMO2, SUMO3 and SUMO4 from the UBLE1A-UBLE1B E1 complex and catalyzes their covalent attachment to other proteins with the help of an E3 ligase such as RANBP2 or CBX4. Necessary for sumoylation of FOXL2 and KAT5. Essential for nuclear architecture and chromosome segregation. -
Tissue specificity
Expressed in heart, skeletal muscle, pancreas, kidney, liver, lung, placenta and brain. Also expressed in testis and thymus. -
Pathway
Protein modification; protein sumoylation. -
Sequence similarities
Belongs to the ubiquitin-conjugating enzyme family. -
Cellular localization
Nucleus. Cytoplasm. Mainly nuclear. In spermatocytes, localizes in synaptonemal complexes. Recruited by BCL11A into the nuclear body. - Information by UniProt
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Database links
- Entrez Gene: 7329 Human
- Entrez Gene: 100044900 Mouse
- Entrez Gene: 22196 Mouse
- Entrez Gene: 25573 Rat
- Omim: 601661 Human
- SwissProt: P63279 Human
- SwissProt: P63280 Mouse
- SwissProt: P63281 Rat
see all -
Alternative names
- C358B7.1 antibody
- p18 antibody
- SUMO 1 protein ligase antibody
see all
Images
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All lanes : Anti-UBE2I / UBC9 antibody [EP2938Y] (ab75854) at 1/10000 dilution
Lane 1 : U937 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HUVEC cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/1000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa -
ab75854, at 1/100 dilution, staining UBE2I / UBC9 in human brain, by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling UBE2I / UBC9 with purified ab75854 at 1/100 dilution (10 µg/mL). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.
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UBE2I / UBC9 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to UBE2I / UBC9 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75854.
Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
Band: 18kDa: UBE2I / UBC9; non specific - 60kDa: We are unsure as to the identity of this extra band. -
Overlay histogram showing HepG2 cells stained with ab75854 (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 (ab75854, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (39)
ab75854 has been referenced in 39 publications.
- Xiao J et al. UBC9 deficiency enhances immunostimulatory macrophage activation and subsequent antitumor T cell response in prostate cancer. J Clin Invest 133:N/A (2023). PubMed: 36626227
- Wang F et al. Novel lncRNA AL033381.2 Promotes Hepatocellular Carcinoma Progression by Upregulating PRKRA Expression. Oxid Med Cell Longev 2022:1125932 (2022). PubMed: 35035655
- Mete B et al. Human immunodeficiency virus type 1 impairs sumoylation. Life Sci Alliance 5:N/A (2022). PubMed: 35181598
- Li F et al. Identification of UBE2I as a Novel Biomarker in ccRCC Based on a Large-Scale CRISPR-Cas9 Screening Database and Immunohistochemistry. Front Mol Biosci 9:813428 (2022). PubMed: 35211510
- Huang M et al. NRIP1 aggravates lung injury caused by Pseudomonas aeruginosa in mice by increasing PIAS1 ubiquitination. Aging (Albany NY) 14:3529-3539 (2022). PubMed: 35460552