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  1. Link

    topoisomerase-ii-alpha--topoisomerase-ii-betatop2b-antibody-epr5377-ab109524.pdf

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Epigenetics and Nuclear Signaling Chromosome Structure Scaffold Proteins
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RecombinantRabMAb

Recombinant Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524)

  • Datasheet
Reviews (2) Submit a question References (14)

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Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
  • Flow Cytometry (Intracellular) - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524)
  • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
  • Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
  • 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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

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Overview

  • 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.

  • Positive control

    • Jurkat, K562, Raw264.7 or PC-12 cell lysate Human breast tissue IF/ICC: HeLa cell line
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR5377
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromosome Structure
    • Scaffold Proteins
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Synthesis
    • Topoisomerases
    • Cancer
    • Drug resistance
    • Topoisomerases

Associated products

  • 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)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
    • K-562 nuclear extract lysate (ab14851)
    • PC-12 nuclear extract lysate (ab14884)

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
ICC/IF
Use a concentration of 10 µg/ml.
WB (2)
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.
Notes
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.
Application notes
Is unsuitable for IP.

Target

  • 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
    • SwissProt: P11388 Human
    • SwissProt: Q02880 Human
    • SwissProt: Q01320 Mouse
    • SwissProt: Q64511 Mouse
    • SwissProt: P41516 Rat
    • Unigene: 156346 Human
    • Unigene: 475733 Human
    • Unigene: 130362 Mouse
    • Unigene: 4237 Mouse
    • Unigene: 90996 Rat
    see all

Images

  • Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
    Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
    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 second


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

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

  • Flow Cytometry (Intracellular) - Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524)
    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) 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
    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.

  • Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
    Western blot - Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377] (ab109524)
    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?

  • Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524)
    Anti-Topoisomerase II alpha + Topoisomerase II beta/TOP2B antibody [EPR5377] (ab109524)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (14)

Publishing research using ab109524? Please let us know so that we can cite the reference in this datasheet.

ab109524 has been referenced in 14 publications.

  • 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
  • Kollárová-Brázdová P  et al. Investigation of Structure-Activity Relationships of Dexrazoxane Analogs Reveals Topoisomerase IIß Interaction as a Prerequisite for Effective Protection against Anthracycline Cardiotoxicity. J Pharmacol Exp Ther 373:402-415 (2020). PubMed: 32253261
  • Schellenberg MJ  et al. Ubiquitin stimulated reversal of topoisomerase 2 DNA-protein crosslinks by TDP2. Nucleic Acids Res 48:6310-6325 (2020). PubMed: 32356875
  • Oury J  et al. MACF1 links Rapsyn to microtubule- and actin-binding proteins to maintain neuromuscular synapses. J Cell Biol 218:1686-1705 (2019). PubMed: 30842214
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-Topoisomerase II alpha + Topoisomerase II beta antibody [EPR5377]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, HT1080)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Treatment
siRNA transfected
Specification
HeLa, HT1080
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jan 05 2017

Western blot abreview for Anti-Topoisomerase II beta antibody [EPR5377]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (4-12 Bis tris)
Sample
Human Cell lysate - whole cell (293T)
Specification
293T
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Jun 20 2014

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