Anti-Topoisomerase II beta/TOP2B antibody (ab125297)
Key features and details
- Rabbit polyclonal to Topoisomerase II beta/TOP2B
- Suitable for: WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Topoisomerase II beta/TOP2B antibody
See all Topoisomerase II beta/TOP2B primary antibodies -
Description
Rabbit polyclonal to Topoisomerase II beta/TOP2B -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Gorilla, Orangutan
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Immunogen
Synthetic peptide corresponding to Human Topoisomerase II beta/TOP2B aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab156303) -
Positive control
- WB: Wild-type NALM-6 cell lysate. HeLa and NIH/3T3 whole cell lysate. Jurkat, K562, MCF7 and HepG2 cell lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab125297 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 WB Use a concentration of 1 µg/ml. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa). Abcam recommends using milk as the blocking agent - 5%Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 183 kDa (predicted molecular weight: 183 kDa). Abcam recommends using milk as the blocking agent - 5%Target
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Function
Control 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 similarities
Belongs to the type II topoisomerase family. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Cytoplasm. Nucleus > nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 282097 Cow
- Entrez Gene: 7155 Human
- Entrez Gene: 21974 Mouse
- Entrez Gene: 361100 Rat
- Omim: 126431 Human
- SwissProt: Q02880 Human
- SwissProt: Q64511 Mouse
- Unigene: 475733 Human
see all -
Alternative names
- Antigen MLAA 44 antibody
- beta isozyme antibody
- DNA topoisomerase 2 beta antibody
see all
Images
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Western blot - Anti-Topoisomerase II beta/TOP2B antibody (ab125297)All lanes : Anti-Topoisomerase II beta/TOP2B antibody (ab125297)
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : TOP2B knockout HEK-293T cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 183 kDa
Observed band size: 183 kDaLanes 1- 2: Merged signal (red and green). Green - ab125297 observed at 183 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab125297 was shown to react with Topoisomerase II beta/TOP2B in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266340 (knockout cell lysate ab257286) was used. Wild-type HEK-293T and TOP2B knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab125297 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-Topoisomerase II beta/TOP2B antibody (ab125297)Lane 1: Wild-type NALM-6 cell lysate (20 µg)
Lane 2: Topoisomerase II beta/TOP2B knockout NALM-6 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: NIH/3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab125297 observed at 200 kDa. Red - loading control, ab7291, observed at 50 kDa.
ab125297 was shown to recognize Topoisomerase II beta/TOP2B when Topoisomerase II beta/TOP2B knockout samples were used, along with additional cross-reactive bands. Wild-type and Topoisomerase II beta/TOP2B knockout samples were subjected to SDS-PAGE. ab125297 and ab7291 (loading control to alpha tubulin) were diluted 1 μg/mL and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Western blot - Anti-Topoisomerase II beta/TOP2B antibody (ab125297)All lanes : Anti-Topoisomerase II beta/TOP2B antibody (ab125297) at 1 µg/ml (Milk blocking - 5%)
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 183 kDa
Observed band size: 183 kDa
Additional bands at: 120 kDa, 150 kDa, 42 kDa, 55 kDa, 66 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes
Abcam recommends using milk as the blocking agent - 5%. This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab125297 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab125297 has been referenced in 2 publications.
- Tadokoro T et al. Mitochondria-dependent ferroptosis plays a pivotal role in doxorubicin cardiotoxicity. JCI Insight 5:N/A (2020). PubMed: 32376803
- Pennington SM et al. Defective protein repair under methionine sulfoxide A deletion drives autophagy and ARE-dependent gene transcription. Redox Biol 16:401-413 (2018). WB . PubMed: 29649787
Images
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Western blot - Anti-Topoisomerase II beta/TOP2B antibody (ab125297)All lanes : Anti-Topoisomerase II beta/TOP2B antibody (ab125297)
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : TOP2B knockout HEK-293T cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 183 kDa
Observed band size: 183 kDaLanes 1- 2: Merged signal (red and green). Green - ab125297 observed at 183 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab125297 was shown to react with Topoisomerase II beta/TOP2B in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266340 (knockout cell lysate ab257286) was used. Wild-type HEK-293T and TOP2B knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab125297 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-Topoisomerase II beta/TOP2B antibody (ab125297)
Lane 1: Wild-type NALM-6 cell lysate (20 µg)
Lane 2: Topoisomerase II beta/TOP2B knockout NALM-6 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: NIH/3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab125297 observed at 200 kDa. Red - loading control, ab7291, observed at 50 kDa.
ab125297 was shown to recognize Topoisomerase II beta/TOP2B when Topoisomerase II beta/TOP2B knockout samples were used, along with additional cross-reactive bands. Wild-type and Topoisomerase II beta/TOP2B knockout samples were subjected to SDS-PAGE. ab125297 and ab7291 (loading control to alpha tubulin) were diluted 1 μg/mL and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Western blot - Anti-Topoisomerase II beta/TOP2B antibody (ab125297)All lanes : Anti-Topoisomerase II beta/TOP2B antibody (ab125297) at 1 µg/ml (Milk blocking - 5%)
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 183 kDa
Observed band size: 183 kDa
Additional bands at: 120 kDa, 150 kDa, 42 kDa, 55 kDa, 66 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes
Abcam recommends using milk as the blocking agent - 5%. This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab125297 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
