Application
Fig1: Western blot analysis of Topoisomerase I on different lysates with Rabbit anti-Topoisomerase I antibody at 1/20,000 dilution. Lane 1: MCF7 cell lysate， Lane 2: Jurkat cell lysate， Lane 3: HepG2 cell lysate， Lane 4: Neuro-2a cell lysate， Lane 5: PC-12 cell lysate， Lysates/proteins at 20 µg/Lane. Predicted band size: 91 kDa Observed band size: 100 kDa Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.Application
Fig2: Immunocytochemistry analysis of MCF7 cells labeling Topoisomerase I with Rabbit anti-Topoisomerase I antibody at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Topoisomerase I antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.Application
Fig3: ICC staining TOP1 in SHG-44 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.Application
Fig4: ICC staining TOP1 in 293 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.Application
Fig5: Immunohistochemical analysis of paraffin-embedded human large intestine tissue with Rabbit anti-Topoisomerase I antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.Application
Fig6: Flow cytometric analysis of HepG2 cells with TOP1 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody| Product Name | Topoisomerase I Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | DNA topoisomerases play essential roles in many DNA metabolic processes including DNA repair. Topoisomerases can introduce DNA damage upon exposure to drugs that stabilize the covalent protein-DNA intermediate of the topoisomerase reaction. Lesions in DNA are also able to trap topoisomerase-DNA intermediates. DNA topoisomerase I (Top1) catalyzes the relaxation of supercoiled DNA by a mechanism of transient DNA strand cleavage characterized by the formation of a phosphotyrosyl bond between the DNA end and active site tyrosine. The antitumor agent camptothecin (CPT) reversibly stabilizes the covalent enzyme-DNA intermediate by inhibiting DNA religation. When a replication fork collides with a DNA Top1 cleavage complex, the covalently bound enzyme must be removed from the DNA 3' end before recombination-dependent replication restart. The tyrosyl-DNA phosphodiesterase Tdp1 and the structure-specific endonuclease Rad1-Rad10 function as primary alternative pathways of Top1 repair in Saccharomyces cerevisiae. In the budding yeast S. cerevisiae, DNA topoisomerases I and II can functionally substitute for each other in removing positive and negative DNA supercoils. |
| Immunogen | recombinant protein |
| Isotype | IgG |
|---|---|
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1,000-1:5,000 ICC:1:50-1:500 IHC:1:50-1:200 FC:1:50-1:100 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | DNA topoisomerase 1 antibody DNA topoisomerase I antibody NUP98 fusion gene antibody TOP 1 antibody TOP I antibody TOP1 antibody TOP1_HUMAN antibody TOPI antibody Topoisomerase (DNA) I antibody Topoisomerase 1 antibody Topoisomerase1 antibody TopoisomeraseI antibody Type I DNA topoisomerase antibody |
|---|---|
| Molecular Weight(MW) | 91 kDa |
| Cellular Localization | Nucleus, nucleoplasm. |
Application
Fig1: Western blot analysis of Topoisomerase I on different lysates with Rabbit anti-Topoisomerase I antibody at 1/20,000 dilution. Lane 1: MCF7 cell lysate， Lane 2: Jurkat cell lysate， Lane 3: HepG2 cell lysate， Lane 4: Neuro-2a cell lysate， Lane 5: PC-12 cell lysate， Lysates/proteins at 20 µg/Lane. Predicted band size: 91 kDa Observed band size: 100 kDa Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
Application
Fig2: Immunocytochemistry analysis of MCF7 cells labeling Topoisomerase I with Rabbit anti-Topoisomerase I antibody at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Topoisomerase I antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
Application
Fig3: ICC staining TOP1 in SHG-44 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig4: ICC staining TOP1 in 293 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig5: Immunohistochemical analysis of paraffin-embedded human large intestine tissue with Rabbit anti-Topoisomerase I antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Application
Fig6: Flow cytometric analysis of HepG2 cells with TOP1 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody| Positive Control | Jurkat, MCF-7, Hela, HepG2, SHG-44, 293, mouse brain tissue. |
|---|---|
| Application Notes | WB:1:1,000-1:5,000 ICC:1:50-1:500 IHC:1:50-1:200 FC:1:50-1:100 |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
