製品の概要

  • 製品名Anti-Topoisomerase I antibody
    Topoisomerase I 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Topoisomerase I
  • アプリケーション適用あり: ICC/IF, WB, IHC-P, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Chicken, Hamster, Cow, Monkey
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Topoisomerase I.

    (Peptide available as ab94391.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following human whole cell lysates: HeLa; Jurkat; HepG2; HEK293; MCF7; Caco2; SHSY5Y; NIH3T3; PC12.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab85038 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
IHC-P Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能The reaction catalyzed by topoisomerases leads to the conversion of one topological isomer of DNA to another.
  • 関連疾患Note=A chromosomal aberration involving TOP1 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with NUP98.
  • 配列類似性Belongs to the eukaryotic type I topoisomerase family.
  • 翻訳後修飾Sumoylated. Lys-117 is the main site of sumoylation. Sumoylation plays a role in partitioning TOP1 between nucleoli and nucleoplasm. Levels are dramatically increased on camptothecin (CPT) treatment.
  • 細胞内局在Nucleus > nucleolus. Nucleus > nucleoplasm. Diffuse nuclear localization with some enrichment in nucleoli. On CPT treatment, cleared from nucleoli into nucleoplasm. Sumolyated forms found in both nucleoplasm and nucleoli.
  • Information by UniProt
  • 参照データベース
  • 別名
    • DNA topoisomerase 1 antibody
    • DNA topoisomerase I antibody
    • NUP98 fusion gene antibody
    • TOP 1 antibody
    • TOP I antibody
    • TOP1 antibody
    • TOP1_HUMAN antibody
    • TOPI antibody
    • Topoisomerase (DNA) I antibody
    • Topoisomerase 1 antibody
    • Topoisomerase1 antibody
    • TopoisomeraseI antibody
    • Type I DNA topoisomerase antibody
    see all

Anti-Topoisomerase I antibody 画像

  • All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 90 kDa
    Observed band size : 90 kDa


    Exposure time : 30 seconds
  • All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 90 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 50 kDa,55 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • Topoisomerase I was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to Topoisomerase I 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, Hek293 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 ab85038.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 90kDa; Topoisomerase I
  • ICC/IF image of ab85038 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85038, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methnaol fixed (5 min) HeLa cells at 1µg/ml.

Anti-Topoisomerase I antibody (ab85038) 使用論文

This product has been referenced in:
  • Guzman-Ayala M  et al. Chd1 is essential for the high transcriptional output and rapid growth of the mouse epiblast. Development 142:118-27 (2015). WB . Read more (PubMed: 25480920) »
  • Boichuk S  et al. Unbiased compound screening identifies unexpected drug sensitivities and novel treatment options for gastrointestinal stromal tumors. Cancer Res 74:1200-13 (2014). Read more (PubMed: 24385214) »

See all 3 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Fibroblast)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 30 µg
Specification Fibroblast
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Feb 01 2017

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