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Anti-Topoisomerase I antibody [EPR5375] (ab109374) is a rabbit monoclonal antibody that is used to detect Topoisomerase I in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 40 publications


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (AB109374), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] (AB109374), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Topoisomerase I antibody [EPR5375] (AB109374), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (AB109374), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (AB109374), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Not recommended
Tested
Tested
Tested
Mouse
Tested
Not recommended
Expected
Expected
Expected
Rat
Tested
Not recommended
Expected
Expected
Expected

Tested
Tested

Species
Mouse
Dilution info
1/100 - 1/250
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species
Rat
Dilution info
1/100 - 1/250
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species
Human
Dilution info
1/100 - 1/250
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/10000 - 1/50000
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500
Notes

For unpurified use at 1/100 - 1/250

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/10 - 1/100
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

Target data

Function

Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(3'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 5'-OH DNA strand. The free DNA strand then rotates around the intact phosphodiester bond on the opposing strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 5'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone (By similarity). Regulates the alternative splicing of tissue factor (F3) pre-mRNA in endothelial cells. Involved in the circadian transcription of the core circadian clock component BMAL1 by altering the chromatin structure around the ROR response elements (ROREs) on the BMAL1 promoter.

Alternative names

Recommended products

Anti-Topoisomerase I antibody [EPR5375] (ab109374) is a rabbit monoclonal antibody that is used to detect Topoisomerase I in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 40 publications

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR5375
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Stable for 12 months at -20°C

Notes

What is this antibody validated in?


Anti-Topoisomerase I antibody [EPR5375] (ab109374) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.

What is the molecular weight of Topoisomerase I?


Anti-Topoisomerase I [EPR5375] (ab109374) specifically detects a band for Topoisomerase I (UniProt: P11387) at a molecular weight of 91kDa.

Trusted by the scientific community


Anti-Topoisomerase I [EPR5375] (ab109374) was first used in a scientific publication in 2011 and has been cited over 40 times in peer-reviewed journals.

Reviewed by scientists


Anti-Topoisomerase I [EPR5375] (ab109374) has over 5 independent reviews from customers.

Trial sizes available!


Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products


We have a range of other formats of antibody clone [EPR5375] also available for your convenience:
ab109374, HRP - HRP Anti-Topoisomerase I antibody [EPR5375] ab196642, Alexa Fluor® 647 - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] ab196930, Alexa Fluor® 488 - Alexa Fluor® 488 Anti-Topoisomerase I antibody [EPR5375] ab197505, Carrier free - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free ab219735



Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Topoisomerase I also known as DNA topoisomerase I or Topo I is an important enzyme in the unwinding of the DNA double helix during processes such as replication and transcription. This enzyme has a molecular mass of approximately 91 kDa and functions by creating transient single-strand breaks in the DNA allowing relaxation of supercoils. Topoisomerase I is widely expressed in both proliferating and non-proliferating cells with higher levels seen in tissues with rapid cell division like the gastrointestinal tract and some immune cells.

Biological function summary

Topoisomerase I plays a significant role in the modulation of DNA topology ensuring proper chromosomal functions during cellular proliferation. It serves as a single polypeptide and does not require a companion for its activity unlike other topoisomerases that may function in complexes. The enzyme’s ability to relieve torsional stress in DNA is essential for maintaining genomic stability and facilitating the smooth progression of the replication fork.

Pathways

Topoisomerase I is vital for DNA replication and transcription processes. It works in concert with other proteins such as helicases and ligases to ensure efficient unwinding and rewinding of DNA strands. The enzyme participates in the DNA damage response pathway where it interacts with proteins like PARP1 to coordinate repair processes. Its action is important in both the S phase of the cell cycle where DNA synthesis occurs and in the G0 phase where cells are in a quiescent state.

Associated diseases and disorders

Topoisomerase I is heavily implicated in oncogenesis with significant associations to colorectal and ovarian cancers. Its heightened activity can lead to genomic instability a hallmark of cancer development. The enzyme also relates to chemotherapeutic resistance where mutations in topoisomerase I lead to reduced drug efficacy. Furthermore inhibitors targeting topoisomerase I such as camptothecin-based drugs exploit its role in cancer aiming to induce DNA damage selectively in rapidly dividing cells. Connections to other proteins like BRCA1 are apparent in these pathways as they both contribute to the DNA repair processes in cells.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling Topoisomerase I with purified ab109374 at 1:100 dilution (1.29 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Topoisomerase I with Purified ab109374 at 1/500 dilution (0.3 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry (Intracellular) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Topoisomerase I with Purified ab109374 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 80% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling Topoisomerase I with purified ab109374 at 1:100 dilution (1.29 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell carcinoma of kidney tissue sections labeling Topoisomerase I with Purified ab109374 at 1:100 dilution (1.29 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

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

    All lanes: Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/50000 dilution

    All lanes: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 91 kDa

    Observed band size: 91 kDa

  • Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    All lanes: Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/10000 dilution

    Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Lane 2: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 20 µg

    Lane 3: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

    Lane 4: Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 91 kDa

    Observed band size: 91 kDa

  • Flow Cytometry (Intracellular) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Overlay histogram showing HepG2 cells stained with ab109374 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109374, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    All lanes: Western blot - Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/10000 dilution

    Lane 1: Jurkat cell lysate at 10 µg

    Lane 2: MCF7 cell lysate at 10 µg

    Lane 3: SW480 cell lysate at 10 µg

    Lane 4: K562 cell lysate at 10 µg

    Predicted band size: 91 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue using ab109374 at a dilution of 1/100. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunohistochemical analysis of paraffin-embedded Human colonic carcinoma tissue using ab109374 at a dilution of 1/100. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] (ab109374), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] (ab109374)

    Immunofluorescent staining of MCF7 cells using ab109374 at a dilution of 1/100.

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Product protocols

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