RabMAb

Anti-UBE2I / UBC9抗体[EP2938Y] - ChIP Grade (ab75854)

概述

性能

应用

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.

应用 Ab评论 说明
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.
ICC 1/100 - 1/250.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

靶标

  • 功能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.
  • 组织特异性Expressed in heart, skeletal muscle, pancreas, kidney, liver, lung, placenta and brain. Also expressed in testis and thymus.
  • 通路Protein modification; protein sumoylation.
  • 序列相似性Belongs to the ubiquitin-conjugating enzyme family.
  • 细胞定位Nucleus. Cytoplasm. Mainly nuclear. In spermatocytes, localizes in synaptonemal complexes. Recruited by BCL11A into the nuclear body.
  • Information by UniProt
  • 数据库链接
  • 别名
    • C358B7.1 antibody
    • p18 antibody
    • SUMO 1 protein ligase antibody
    • SUMO conjugating enzyme UBC9 antibody
    • SUMO-conjugating enzyme UBC9 antibody
    • SUMO-protein ligase antibody
    • SUMO1 protein ligase antibody
    • UBC9 antibody
    • UBC9_HUMAN antibody
    • UBCE9 antibody
    • Ube2i antibody
    • Ubiquitin carrier protein 9 antibody
    • Ubiquitin carrier protein antibody
    • Ubiquitin carrier protein I antibody
    • Ubiquitin conjugating enzyme 9 antibody
    • Ubiquitin conjugating enzyme E2I (homologous to yeast UBC9) antibody
    • Ubiquitin conjugating enzyme E2I (UBC9 homolog, yeast) antibody
    • Ubiquitin conjugating enzyme UbcE2A antibody
    • Ubiquitin like protein SUMO 1 conjugating enzyme antibody
    • Ubiquitin protein ligase E2I antibody
    • Ubiquitin-conjugating enzyme E2 I antibody
    • Ubiquitin-protein ligase I antibody
    see all

Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade 图像

  • All lanes : Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade (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
    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.
  • 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.

Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade (ab75854)参考文献

This product has been referenced in:
  • Hsieh YL  et al. Ubc9 acetylation modulates distinct SUMO target modification and hypoxia response. EMBO J 32:791-804 (2013). Read more (PubMed: 23395904) »
  • Santos A  et al. SUMOylation affects the interferon blocking activity of the influenza A nonstructural protein NS1 without affecting its stability or cellular localization. J Virol 87:5602-20 (2013). Read more (PubMed: 23468495) »

See all 10 Publications for this product

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6
Sample Mouse Tissue sections (Spinal cord 16 micron cryosection)
Specification Spinal cord 16 micron cryosection
Permeabilization Yes - 0.5% Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 08 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Hela)
Total protein in input 100 µg
Specification Hela
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

提交于 Aug 27 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (LIVER)
Specification LIVER
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37%FORMALDEHYDE
Detection step Semiquantitative PCR
Positive control POL11
Negative control IgG
Username

Mr. Natarajan Balasubramaniyan

Verified customer

提交于 May 19 2012

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"