The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
Use a concentration of 1 µg/ml.
Use a concentration of 5 µg/ml.
FunctionThe reaction catalyzed by topoisomerases leads to the conversion of one topological isomer of DNA to another.
Involvement in diseaseNote=A chromosomal aberration involving TOP1 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with NUP98.
Sequence similaritiesBelongs 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 localizationNucleus > 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.
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 - 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.
References for Anti-Topoisomerase I antibody (ab85038)
This product has been referenced in:
Guzman-Ayala M et al. Chd1 is essential for the high transcriptional output and rapid growth of the mouse epiblast. Development142:118-27 (2015).
Read more (PubMed: 25480920) »
Boichuk S et al. Unbiased compound screening identifies unexpected drug sensitivities and novel treatment options for gastrointestinal stromal tumors. Cancer Res74:1200-13 (2014).
Read more (PubMed: 24385214) »