Recombinant Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5377] to Topoisomerase II alpha + Topoisomerase II beta/TOP2B
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377]
See all Topoisomerase II alpha+Topoisomerase II beta primary antibodies -
Description
Rabbit monoclonal [EPR5377] to Topoisomerase II alpha + Topoisomerase II beta/TOP2B -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Jurkat, K562, Raw264.7 or PC-12 cell lysate Human breast tissue IF/ICC: HeLa cell line
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5377 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 647 Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab200993)
- Alexa Fluor® 488 Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab201504)
- Alexa Fluor® 555 Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab206605)
- Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] - BSA and Azide free (ab239984)
- Alexa Fluor® 568 Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab312562)
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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ICC/IF |
Use a concentration of 10 µg/ml.
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WB | (2) |
1/10000 - 1/50000. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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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 (Intra)
Use at an assay dependent concentration. |
Target
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Cellular localization
Topoisomerase II alpha: Cytoplasm. Nucleus > nucleoplasm. Generally located in the nucleoplasm. Topoisomerase II beta: Cytoplasm. Nucleus > nucleolus. -
Database links
- Entrez Gene: 7153 Human
- Entrez Gene: 7155 Human
- Entrez Gene: 21973 Mouse
- Entrez Gene: 21974 Mouse
- Entrez Gene: 360243 Rat
- Entrez Gene: 361100 Rat
- Omim: 126430 Human
- Omim: 126431 Human
see all
Images
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All lanes : 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 : 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 secondBlocking and diluting buffer and concentration: 5% NFDM/TBST.
TOP II alpha recombinant protein (aa 1-220) containing a his-tag.
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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)
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Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase II alpha + 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha + Topoisomerase II beta 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.
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All lanes : Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B 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?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (22)
ab109524 has been referenced in 22 publications.
- Ke F et al. Replication and transcription machinery for ranaviruses: components, correlation, and functional architecture. Cell Biosci 12:6 (2022). PubMed: 34991685
- Flörkemeier I et al. High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer. Cancers (Basel) 14:N/A (2021). PubMed: 35008166
- Mengoli V et al. Deprotection of centromeric cohesin at meiosis II requires APC/C activity but not kinetochore tension. EMBO J 40:e106812 (2021). PubMed: 33644894
- Modesto PN et al. Green Tea (Camellia sinensis) Extract Increased Topoisomerase IIß, Improved Antioxidant Defense, and Attenuated Cardiac Remodeling in an Acute Doxorubicin Toxicity Model. Oxid Med Cell Longev 2021:8898919 (2021). PubMed: 34035878
- Huang ZG et al. MiRNA-145-5p expression and prospective molecular mechanisms in the metastasis of prostate cancer. IET Syst Biol 15:1-13 (2021). PubMed: 33527765