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RabMAb

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

製品の概要

  • 製品名
    Anti-UBE2I / UBC9 antibody [EP2938Y] - ChIP Grade
    UBE2I / UBC9 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP2938Y] to UBE2I / UBC9 - ChIP Grade
  • アプリケーション
    適用あり: ChIP, WB, IP, IHC-P, ICC, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues near the N-terminus of human UBE2I/ UBC9

  • ポジティブ・コントロール
    • U937, HeLa, Jurkat or HUVEC cell lysates. Human brain tissue.
  • 特記事項

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    Expressed in heart, skeletal muscle, pancreas, kidney, liver, lung, placenta and brain. Also expressed in testis and thymus.
  • パスウェイ
    Protein modification; protein sumoylation.
  • 配列類似性
    Belongs to the ubiquitin-conjugating enzyme family.
  • 細胞内局在
    Nucleus. Cytoplasm. Mainly nuclear. In spermatocytes, localizes in synaptonemal complexes. Recruited by BCL11A into the nuclear body.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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 画像

  • 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) 使用論文

This product has been referenced in:
  • He X  et al. Characterization of the loss of SUMO pathway function on cancer cells and tumor proliferation. PLoS One 10:e0123882 (2015). Read more (PubMed: 25860128) »
  • Balasubramaniyan N  et al. SUMOylation of the farnesoid X receptor (FXR) regulates the expression of FXR target genes. J Biol Chem 288:13850-62 (2013). Read more (PubMed: 23546875) »

See all 13 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

投稿 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

投稿 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

投稿 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"

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