Anti-Topoisomerase I antibody (ab85038)
Key features and details
- Rabbit polyclonal to Topoisomerase I
- Suitable for: ICC/IF, WB, IP
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-Topoisomerase I antibody
See all Topoisomerase I primary antibodies -
Description
Rabbit polyclonal to Topoisomerase I -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Hamster, Cow, Monkey
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Immunogen
Synthetic peptide corresponding to Human Topoisomerase I aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab94391) -
General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab85038 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 ICC/IF Use a concentration of 1 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa). IP Use a concentration of 5 µg/ml. 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: 534799 Cow
- Entrez Gene: 7150 Human
- Entrez Gene: 21969 Mouse
- Entrez Gene: 64550 Rat
- Omim: 126420 Human
- SwissProt: P11387 Human
- SwissProt: Q04750 Mouse
- SwissProt: Q9WUL0 Rat
see all -
Alternative names
- DNA topoisomerase 1 antibody
- DNA topoisomerase I antibody
- NUP98 fusion gene antibody
see all
Images
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Western blot - Anti-Topoisomerase I antibody (ab85038)All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 90 kDa
Observed band size: 90 kDa
Exposure time: 30 seconds
Human DNA topoisomerase 1 contains a potential sumoylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted. -
Western blot - Anti-Topoisomerase I antibody (ab85038)All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 90 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa, 55 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes -
Immunoprecipitation - Anti-Topoisomerase I antibody (ab85038)Topoisomerase I was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to Topoisomerase I and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85038.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 90kDa; Topoisomerase I -
Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody (ab85038)ICC/IF image of ab85038 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85038, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methnaol fixed (5 min) HeLa cells at 1µg/ml.
Protocols
References (5)
ab85038 has been referenced in 5 publications.
- Lv C et al. Analysis of topoisomerase I expression and identification of predictive markers for efficacy of topotecan chemotherapy in small cell lung cancer. Thorac Cancer 9:1166-1173 (2018). PubMed: 30058109
- Bansal K et al. The transcriptional regulator Aire binds to and activates super-enhancers. Nat Immunol 18:263-273 (2017). PubMed: 28135252
- Guzman-Ayala M et al. Chd1 is essential for the high transcriptional output and rapid growth of the mouse epiblast. Development 142:118-27 (2015). WB . PubMed: 25480920
- Boichuk S et al. Unbiased compound screening identifies unexpected drug sensitivities and novel treatment options for gastrointestinal stromal tumors. Cancer Res 74:1200-13 (2014). PubMed: 24385214
- Qin H et al. Transcriptional analysis of pluripotency reveals the Hippo pathway as a barrier to reprogramming. Hum Mol Genet 21:2054-67 (2012). WB . PubMed: 22286172
Images
-
Western blot - Anti-Topoisomerase I antibody (ab85038)All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 90 kDa
Observed band size: 90 kDa
Exposure time: 30 seconds
Human DNA topoisomerase 1 contains a potential sumoylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted. -
Western blot - Anti-Topoisomerase I antibody (ab85038)All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 90 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa, 55 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
-
Immunoprecipitation - Anti-Topoisomerase I antibody (ab85038)Topoisomerase I was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to Topoisomerase I and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85038.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 90kDa; Topoisomerase I -
Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody (ab85038)ICC/IF image of ab85038 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85038, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methnaol fixed (5 min) HeLa cells at 1µg/ml.
