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Anti-UBE2I Antibody EP2938Y
Also for UBE2I (NM_194259)
|A synthetic peptide corresponding to residues near the N-terminus of human UBC9 was used as an immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IP, FC
||ChIP: Use at an assay dependent concentration; WB: 1:1000 - 1:10000; IP: Use at an assay dependent concentration; IHC-P: 1:100 - 1:250; ICC: 1:100 - 1:250; FC: 1:50
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Homo sapiens ubiquitin-conjugating enzyme E2I (UBE2I), transcript variant 2|
|C358B7.1; P18; UBC9|
|UBCs are a family of proteins directly involved in ubiquitination of proteins and in the control of cellular processes through the targeting of key regulatory proteins for degradation UBC9 is a homologue of the class E2 ubiquitin-conjugating enzymes (UBCs) (1). UBC9 possesses a distinct electrostatic potential distribution that may provide possible clues to its ability to interact with other proteins. It was suggested that different UBCs may utilize catalytic mechanisms of varying efficiency and substrate specificity (2). UBC9 has been shown to catalyze conjugation of a small ubiquitin-like molecule-1 (SUMO-1) to a variety of target proteins. SUMO-1 modifications were implicated in the targeting of proteins to the nuclear envelope and certain intranuclear structures and in converting proteins resistant to ubiquitin-mediated degradation. UBC9 also interacts with the androgen receptor (AR), a member of the steroid receptor family of ligand-activated transcription factors. It has the ability to act as an AR co-regulator in a fashion independent of its ability to catalyze SUMO-1 conjugation. (3). |
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Western blot - UBE2I / UBC9 antibody [EP2938Y]; All lanes : Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade 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.Goat anti-rabbit HRP at 1/1000 dilution.Predicted band size : 18 kDa.Observed band size : 18 kDa.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - UBE2I / UBC9 antibody [EP2938Y]; ab75854, at 1/100 dilution, staining UBE2I / UBC9 in human brain, by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.
Immunohistochemistry (Frozen sections) - Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade; 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 ug/ml and an AlexaFluor?700-conjugated goat anti-rabbit IgG (1 ug/ml) was used as the secondary antibody.
Immunoprecipitation - Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade; 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.
Flow Cytometry-Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade(ab75854); 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 for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed.