Anti-Topoisomerase II alpha 抗体 [EP1102Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Topoisomerase II alpha with ab52934 at a dilution of 1/8000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Paraffin embedded human testis tissue slides were dewaxed, followed by heat mediated antigen retrieval with Citrate buffer (pH 6.0, Epitope Retrieval ER1 Solution) for 20 mins. Endogenous peroxidase activity was blocked by Refine Detection Kit Peroxide Block for 10 mins. The sections were then incubated with ab52934 (1/4500) at room temperature for 30 mins, followed by a ready to use LeicaDS9800 (BOND Polymer Refine Detection kit). The sections were counterstained with Hematoxylin. The images were taken with a Leica Aperio AT2.

ab52934 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in IHC-P. The image shows the staining intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Topoisomerase II alpha with ab52934 at a dilution of 1/8000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse colon tissue labelling Topoisomerase II alpha with ab52934 at a dilution of 1/8000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat colon tissue labelling Topoisomerase II alpha with ab52934 at a dilution of 1/8000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue labelling Topoisomerase II alpha with ab52934 at a dilution of 1/8000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue labelling Topoisomerase II alpha with ab52934 at a dilution of 1/8000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Lab

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of HeLa whole cell lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab52934 (1/1000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/20,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique.

ab52934 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant for temperatures ranging between 4°C, 22°C and 37°, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

All lanes:

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (ab52934) at 1/1000 dilution

All lanes:

HeLa whole cell lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

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

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Exposure time: 40s

• WB

Lab

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Exposure time :

Lanes 1-3 : 1 second.
Lanes 4-7 : 3 minutes.
Lane 8 : 7 seconds.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (ab52934) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Jurkat (human acute T cell leukemia) whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma) whole cell lysate at 20 µg

Lane 4:

Human breast cancer lysate at 20 µg

Lane 5:

Mouse thymus lysate at 20 µg

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 7:

Rat thymus lysate at 20 µg

Lane 8:

Rat testis lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 174 kDa

Observed band size: 174 kDa

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• WB

Lab

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

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

All lysate were prepared using 1%SDS Hot lysis method.

We recommend to use 1%SDS Hot lysis prepare method to get desired Western Blot results.

All lanes:

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (ab52934) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method 20μg at 20 µg

Secondary

All lanes:

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

Predicted band size: 174 kDa

false

Exposure time: 7s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Western Blotting using Anti-Topoisomerase II alpha antibody [EP1102Y], ab52934. Publication image from Hakem, R. et al., 2016, Nat Commun, 27558965. Legend direct from paper.

RNF168 interacts with TOP2α.(a) Identification of RNF168-associated proteins. A representative SDS–polyacrylamide gel electrophoresis of Flag-RNF168-associated proteins. Flag-tagged RNF168 was transfected in HEK293T cells and pull-down analysis was performed 48 h later. Protein bands were detected by silver staining. Protein bands were identified by mass spectrometry analysis following in-gel protease digestion. (b) HEK293T cells were transfected as indicated with HA-tagged RNF168 and Flag-TOP2α expression vectors. Cells were lysed and IP was performed using anti-Flag antibody. The resulting precipitates were subjected to IB analysis with the indicated antibodies. WCL, whole-cell lysate. (c) TOP2α, RNF168 and IgG (control) immunoprecipitates from HEK293T cells were examined by IB as indicated. (b,c) Data are representative of three independent experiments. (d) Cells treated with EdU were used for detection of localization patterns of TOP2α (Alexa Fluor 488) and RNF168 (Alexa Fluor 594) using confocal microscopy. Scale bar, 20 µm.

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• WB

CiteAb

Western blot - Anti-Topoisomerase II alpha antibody [EP1102Y] (AB52934)

Western Blotting using Anti-Topoisomerase II alpha antibody [EP1102Y], ab52934. Publication image from Hakem, R. et al., 2016, Nat Commun, 27558965. Legend direct from paper.

RNF168 mediates ubiquitylation of TOP2α.(a) RIDDLE cells reconstituted with HA-RNF168 or HA-empty vector, and control HA-RNF168-reconstituted RIDDLE cells with TOP2α knock down were lysed and WCL subjected to IP with anti-TOP2α or IgG (control) antibodies. Immunoprecipitates were blotted with the indicated antibodies to detect ubiquitylated TOP2α. (b) Human breast cancer cell lines T47D and MDA-MB-231 knocked down for RNF168 (Sh.RNF168) and their control expressing ShRNA control (sh.Ctr) were examined for their level of ubiquitylated TOP2α as in a. (c) Rnf168−/−, Brca1−/− and WT MEFs were lysed and subjected to IP with anti-Top2α or IgG (control) antibodies. IPs from WCL were blotted with the indicated antibodies. (d) HEK293T cells were transfected with RNF168 (WT or mutant Rnf168-C21S), Flag-TOP2α and HA-Ub vectors as indicated. WCL were subjected to IP with anti-Flag, and IB analysis was performed using anti-HA antibody to detect ubiquitylated Flag-TOP2α. (e) Nuclear extracts from RIDDLE cells reconstituted with HA-RNF168 or HA-empty vector were subjected to IP with anti-TOP2α or IgG (control) antibodies. Immunoprecipitates were blotted with the indicated antibodies against K63- and K48-Ub linkages. (f) In vitro ubiquitylation of recombinant TOP2α in the presence of recombinant RNF168 (500 ng for lane 5, 1 µg for lanes 2, 3 and 6 and 2 µg for lane7), UBE1 (E1), UBE2E2 (E2) and Ub. Nuc, nuclear extract; WCL, whole-cell lysate.

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