Recombinant Anti-Topoisomerase I antibody [EPR5375] (ab109374)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5375] to Topoisomerase I
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Topoisomerase I antibody [EPR5375]
See all Topoisomerase I primary antibodies -
Description
Rabbit monoclonal [EPR5375] to Topoisomerase I -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB; Jurkat, SW480, MCF7, HepG2, Neuro-2a, PC-12, and K562 cell lysates; IHC-P: Human breast carcinoma, Human colonic carcinoma tissues, Human clear cell carcinoma of kidney and Mouse kidney, and Rat colon tissue; ICC/IF: MCF7 cells;
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5375 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109374 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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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (5) |
1/10000 - 1/50000. Predicted molecular weight: 91 kDa.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/500.
For unpurified use at 1/100 - 1/250 |
Notes |
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Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Predicted molecular weight: 91 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/500. For unpurified use at 1/100 - 1/250 |
Target
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Function
The reaction catalyzed by topoisomerases leads to the conversion of one topological isomer of DNA to another. -
Involvement in disease
Note=A chromosomal aberration involving TOP1 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with NUP98. -
Sequence similarities
Belongs to the eukaryotic type I topoisomerase family. -
Post-translational
modificationsSumoylated. Lys-117 is the main site of sumoylation. Sumoylation plays a role in partitioning TOP1 between nucleoli and nucleoplasm. Levels are dramatically increased on camptothecin (CPT) treatment. -
Cellular localization
Nucleus > nucleolus. Nucleus > nucleoplasm. Diffuse nuclear localization with some enrichment in nucleoli. On CPT treatment, cleared from nucleoli into nucleoplasm. Sumolyated forms found in both nucleoplasm and nucleoli. - Information by UniProt
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Database links
- Entrez Gene: 7150 Human
- Entrez Gene: 21969 Mouse
- Entrez Gene: 64550 Rat
- Omim: 126420 Human
- SwissProt: P11387 Human
- SwissProt: Q04750 Mouse
- SwissProt: Q9WUL0 Rat
- Unigene: 472737 Human
see all -
Alternative names
- DNA topoisomerase 1 antibody
- DNA topoisomerase I antibody
- NUP98 fusion gene antibody
see all
Images
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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 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.
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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 Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (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.
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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, 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).
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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 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.
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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 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.
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Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/50000 dilution (Purified) + PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 91 kDa
Observed band size: 91 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/10000 dilution (Purified)
Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 4 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 91 kDa
Observed band size: 91 kDa -
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) (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.
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All lanes : Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/10000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : SW480 cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 91 kDa -
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.
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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.
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Immunofluorescent staining of MCF7 cells using ab109374 at a dilution of 1/100.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (36)
ab109374 has been referenced in 36 publications.
- Beghè C & Gromak N R-Loop Immunoprecipitation: A Method to Detect R-Loop Interacting Factors. Methods Mol Biol 2528:215-237 (2022). PubMed: 35704194
- Ding X et al. PARP1-SNAI2 transcription axis drives resistance to PARP inhibitor, Talazoparib. Sci Rep 12:12501 (2022). PubMed: 35864202
- Kuzin V et al. TOP1 CAD-seq: A protocol to map catalytically engaged topoisomerase 1 in human cells. STAR Protoc 3:101581 (2022). PubMed: 35942340
- Perry M et al. USP11 mediates repair of DNA-protein cross-links by deubiquitinating SPRTN metalloprotease. J Biol Chem 296:100396 (2021). PubMed: 33567341
- Oliveira ÉA et al. TOP1 modulation during melanoma progression and in adaptative resistance to BRAF and MEK inhibitors. Pharmacol Res 173:105911 (2021). PubMed: 34560251