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AB314011

Anti-Topoisomerase I antibody [6D8]

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Rabbit Monoclonal Topoisomerase I antibody. Suitable for WB, IHC-P, IP, Flow Cyt and reacts with Human samples. Immunogen corresponding to Synthetic Peptide within Human TOP1.

View Alternative Names

DNA topoisomerase 1, DNA topoisomerase I, TOP1

5 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [6D8] (AB314011)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [6D8] (AB314011)

Immunohistochemical analysis of ab314011 diluted at 1 : 100 and staining in paraffin-embedded human colon cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.

Flow Cytometry - Anti-Topoisomerase I antibody [6D8] (AB314011)
  • Flow Cyt

Supplier Data

Flow Cytometry - Anti-Topoisomerase I antibody [6D8] (AB314011)

Overlay histogram showing HepG2 (Human liver hepatocellular carcinoma cell line) cells stained with ab314011 (pink line) at 1 : 50. The cells were fixed with 70% Ethylalcohol (18h) and then incubated in 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1μg/1x10E6 cells) for 1 h at 4°C. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 30min at 4°C. Control antibody (green line) was Rabbit IgG (1μg/1x10E6 cells) used under the same conditions. Acquisition of >10,000 events was performed.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [6D8] (AB314011)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [6D8] (AB314011)

Immunohistochemical analysis of ab314011 diluted at 1 : 100 and staining in paraffin-embedded human small intestine tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.

Immunoprecipitation - Anti-Topoisomerase I antibody [6D8] (AB314011)
  • IP

Supplier Data

Immunoprecipitation - Anti-Topoisomerase I antibody [6D8] (AB314011)

Immunoprecipitating Topoisomerase I in K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate Lane 1 : Rabbit control IgG instead of ab314011 in K562 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000) Lane 2 : ab314011 (2µg)+ K562 whole cell lysate (500µg) Lane 3 : K562 whole cell lysate (10µg)

All lanes:

Immunoprecipitation - Anti-Topoisomerase I antibody [6D8] (ab314011) at 2 µg

Predicted band size: 91 kDa

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Western blot - Anti-Topoisomerase I antibody [6D8] (AB314011)
  • WB

Supplier Data

Western blot - Anti-Topoisomerase I antibody [6D8] (AB314011)

All lanes:

Western blot - Anti-Topoisomerase I antibody [6D8] (ab314011) at 1/2000 dilution

Lane 1:

HeLa whole cell lysate

Lane 2:

MCF7 whole cell lysate

Lane 3:

K562 whole cell lysate

Lane 4:

HL-60 whole cell lysate

Lane 5:

PC-3 whole cell lysate

Secondary

All lanes:

Goat polyclonal to rabbit IgG at 1/50000 dilution

Observed band size: 91 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

6D8

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

IP, WB, Flow Cyt, IHC-P

applications

Immunogen

Synthetic Peptide within Human TOP1.

P11387

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/500 - 1/5000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/50 - 1/200", "IHCP-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/200 - 1/1000", "IP-species-notes": "<p></p>", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "1/20 - 1/200", "FlowCyt-species-notes": "<p></p>" } } }
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Product details

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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.88% Sodium chloride
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
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Supplementary information

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

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.

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.
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Product protocols

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

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.
See full target information TOP1
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Product promise

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For full details, please see our Terms & Conditions

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