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
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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
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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 < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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 | (1) |
Use a concentration of 1 µg/ml.
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| WB | (1) |
Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
|
| IP | (1) |
Use a concentration of 5 µg/ml.
|
| Notes |
|---|
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ICC/IF
Use a concentration of 1 µg/ml. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa). |
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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
Datasheets and documents
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SDS download
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Datasheet download
References (6)
ab85038 has been referenced in 6 publications.
- Hazan I et al. Activation of Oncogenic Super-Enhancers Is Coupled with DNA Repair by RAD51. Cell Rep 29:560-572.e4 (2019). PubMed: 31618627
- 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
Customer reviews and Q&As
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