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Rabbit Recombinant Monoclonal Topoisomerase I antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

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Images

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] (AB196930), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] (AB196930), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Storage buffer

pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% 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
ICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Mouse
Not recommended
Not recommended
Rat
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/100

Notes

This product gave a positive signal in T47D cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

This product gave a positive signal in T47D cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

ab199093 - Rabbit monoclonal IgG (Alexa Fluor®; 647), is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

ab199093 - Rabbit monoclonal IgG (Alexa Fluor®; 647), is suitable for use as an isotype control with this antibody.

Species

Mouse

Dilution info

-

Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

3 products for Alternative Version

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 ARNTL/BMAL1 by altering the chromatin structure around the ROR response elements (ROREs) on the ARNTL/BMAL1 promoter.

Alternative names

DNA topoisomerase 1, DNA topoisomerase I, TOP1

Recommended products

Rabbit Recombinant Monoclonal Topoisomerase I antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

Alternative names

DNA topoisomerase 1, DNA topoisomerase I, TOP1

Key facts

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

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

Avoid freeze / thaw cycle, Store in the dark

Notes

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

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.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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.

2 product images

  • Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] (ab196930), expandable thumbnail

    Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] (ab196930)

    Overlay histogram showing T47D cells stained with ab196930 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab196930, 1/500 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

    This antibody gave a positive signal in T47D fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] (ab196930), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Topoisomerase I antibody [EPR5375] (ab196930)

    ab196930 staining Topoisomerase I in T47D cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196930 at a 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com