Recombinant
RabMAb

Recombinant Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)

Overview

  • Product name

    Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377]
    See all Topoisomerase II alpha+Topoisomerase II beta primary antibodies
  • Description

    Rabbit monoclonal [EPR5377] to Topoisomerase II alpha + Topoisomerase II beta
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Topoisomerase II beta aa 50-150. The exact sequence is proprietary.
    Database link: Q02880

  • Positive control

    • Jurkat, K562, Raw264.7 or PC-12 cell lysate Human breast tissue IF/ICC: HeLa cell line
  • General notes

     

     

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab109524 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 10 µg/ml.
WB 1/10000 - 1/50000. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • Target

    Images

    • All lanes : Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524) at 1/1000 dilution

      Lane 1 : ab109524 on TOP II alpha recombinant protein 10ng
      Lane 2 : Anti-His rabbit serum on TOP II alpha recombinant protein 10ng

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 183 kDa
      Observed band size: 24 kDa
      why is the actual band size different from the predicted?


      Exposure time: 1 second


      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      TOP II alpha recombinant protein (aa 1-220) containing a his-tag.

    • ab109524 staining Topoisomerase II alpha + Topoisomerase II beta in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black)

      Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    • ICC/IF image of ab109524 stained HeLa cells. The cells were 4% Formaldehyde 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 (ab109524, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

    • ab109524, at a 1/100 dilution, staining Human Topoisomerase II alpha + Topoisomerase II beta in breast tissue, using Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • All lanes : Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524) at 1/50000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : Raw264.7 cell lysate
      Lane 4 : PC-12 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 183 kDa
      Observed band size: 180 kDa why is the actual band size different from the predicted?

    References

    This product has been referenced in:

    • Liu LM  et al. DNA topoisomerase 1 and 2A function as oncogenes in liver cancer and may be direct targets of nitidine chloride. Int J Oncol 53:1897-1912 (2018). Read more (PubMed: 30132517) »
    • Wette SG  et al. Characterization of muscle ankyrin repeat proteins in human skeletal muscle. Am J Physiol Cell Physiol 313:C327-C339 (2017). Read more (PubMed: 28615162) »
    See all 6 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa, HT1080)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Treatment
    siRNA transfected
    Specification
    HeLa, HT1080
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jan 05 2017

    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (4-12 Bis tris)
    Sample
    Human Cell lysate - whole cell (293T)
    Specification
    293T
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Jun 20 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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