Product nameAnti-UBE2I / UBC9 antibody [EP2938Y]
See all UBE2I / UBC9 primary antibodies
DescriptionRabbit monoclonal [EP2938Y] to UBE2I / UBC9
Tested applicationsSuitable for: IHC-Fr, ChIP, WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human UBE2I/ UBC9 aa 1-100 (N terminal). The exact sequence is proprietary.
- U937, HeLa, Jurkat or HUVEC cell lysates. Human brain tissue. ICC/IF: Jurkat cells
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.
Storage instructionsShipped 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 bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.5% BSA
Concentration information loading...
PurityProtein A purified
ChIP Related Products
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.
|IHC-Fr||Use a concentration of 1 µg/ml. PubMed: 22016779|
|ChIP||Use at an assay dependent concentration.|
|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.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionAccepts 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 specificityExpressed in heart, skeletal muscle, pancreas, kidney, liver, lung, placenta and brain. Also expressed in testis and thymus.
PathwayProtein modification; protein sumoylation.
Sequence similaritiesBelongs to the ubiquitin-conjugating enzyme family.
Cellular localizationNucleus. Cytoplasm. Mainly nuclear. In spermatocytes, localizes in synaptonemal complexes. Recruited by BCL11A into the nuclear body.
- Information by UniProt
- C358B7.1 antibody
- p18 antibody
- SUMO 1 protein ligase antibody
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.
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.
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.
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.
ab75854 staining UBE2I / UBC9 (red) in cerebral cortex of Ubc9 transgenic mice, by Immunohistochemistry (Frozen sections). Left panel shows all layers of cerebral cortex, and right panel shows an enlarged region of the Layer III-External pyramidal area.
Samples were incubated with primary antibody at 1 µg/ml and an AlexaFluor®700-conjugated goat anti-rabbit IgG (1 µg/ml) was used as the secondary antibody.
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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This product has been referenced in:
- Gao K et al. SUMO peptidase ULP-4 regulates mitochondrial UPR-mediated innate immunity and lifespan extension. Elife 8:N/A (2019). Read more (PubMed: 30642431) »
- Carmichael RE et al. Insulin-dependent GLUT4 trafficking is not regulated by protein SUMOylation in L6 myocytes. Sci Rep 9:6477 (2019). Read more (PubMed: 31019221) »