The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
1/10000 - 1/50000. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notesIs unsuitable for Flow Cyt or IP.
FunctionControl of topological states of DNA by transient breakage and subsequent rejoining of DNA strands. Topoisomerase II makes double-strand breaks. Indirectly ivolved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene.
Sequence similaritiesBelongs to the type II topoisomerase family.
Post-translational modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Anti-Topoisomerase II beta antibody [EPR5377] images
Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase II beta antibody [EPR5377] (ab109524)
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.
Western blot - Topoisomerase II beta antibody [EPR5377] (ab109524)
All lanes : Anti-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