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.
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) 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) 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) 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 -
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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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