Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (AB109524)

ab109524 staining Topoisomerase II alpha + Topoisomerase II beta in the human cell line HeLa (human cervix adenocarcinoma) by intracellular 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 Fluorr^® 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)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (AB109524)

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.

• WB

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Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (AB109524)

All lanes:

Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524) at 1/50000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

Raw264.7 cell lysate at 10 µg

Lane 4:

PC-12 cell lysate at 10 µg

Observed band size: 180 kDa

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• WB

Lab

Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (AB109524)

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

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

All lanes:

Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B 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:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 24 kDa

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Exposure time: 1s