Product nameAnti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377]
See all Topoisomerase II alpha+Topoisomerase II beta primary antibodies
DescriptionRabbit monoclonal [EPR5377] to Topoisomerase II alpha + Topoisomerase II beta
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
Unsuitable for: IP
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Topoisomerase II beta aa 50-150. The exact sequence is proprietary.
Database link: Q02880
- Jurkat, K562, Raw264.7 or PC-12 cell lysate Human breast tissue IF/ICC: HeLa cell line
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
- Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (Alexa Fluor® 647) (ab200993)
- Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (Alexa Fluor® 488) (ab201504)
- Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (Alexa Fluor® 555) (ab206605)
- Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] - BSA and Azide free (ab239984)
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.
|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.|
Cellular localizationTopoisomerase II alpha: Cytoplasm. Nucleus > nucleoplasm. Generally located in the nucleoplasm. Topoisomerase II beta: Cytoplasm. Nucleus > nucleolus.
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
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?
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) »