Topoisomerase I Recombinant Rabbit Monoclonal Antibody [SC69-03] - BSA and Azide free
Catalog# HA750227
Topoisomerase I Recombinant Rabbit Monoclonal Antibody [SC69-03] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Topoisomerase I Recombinant Rabbit Monoclonal Antibody [SC69-03] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human TOP1 aa 1-49 / 765.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC
Target Molecular Weight
Predicted band size: 91 kDa
Positive Control
MCF7 cell lysate, Jurkat cell lysate, HepG2 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, MCF7, Hela, SHG-44, 293, HepG2, mouse brain tissue, human large intestine tissue, mouse hippocampus tissue.
Conjugation
unconjugated
Clone Number
SC69-03
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
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.
Background References
1. Groh M et al. R-loops associated with triplet repeat expansions promote gene silencing in Friedreich ataxia and fragile X syndrome. PLoS Genet 10:e1004318 (2014).
2. Bhatia V et al. BRCA2 prevents R-loop accumulation and associates with TREX-2 mRNA export factor PCID2. Nature 511:362-5 (2014).
Sequence Similarity
Belongs to the type IB topoisomerase family.
Tissue Specificity
Endothelial cells.
Post-translational Modification
Sumoylated. 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.; Phosphorylation at Ser-506 by CK2 increases binding to supercoiled DNA and sensitivity to camptothecin.
Subcellular Location
Nucleus, nucleoplasm.
Synonyms
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
ExpandImages
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This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Western blot analysis of Topoisomerase I on different lysates with Rabbit anti-Topoisomerase I antibody (ET1610-62) 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 (ET1610-62) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of MCF7 cells labeling Topoisomerase I with Rabbit anti-Topoisomerase I antibody (ET1610-62) 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 (ET1610-62) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
ICC staining of Topoisomerase I in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
ICC staining of Topoisomerase I in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
ICC staining of Topoisomerase I in 293 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of Topoisomerase I was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-62, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human large intestine tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1610-62) 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. -
This data was developed using ET1610-62, the same antibody clone in a different buffer formulation.
"Immunocytochemistry analysis of Neuro-2a cells labeling Topoisomerase I with Rabbit anti-Topoisomerase I antibody (ET1610-62) at 1/500 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 (ET1610-62) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution."
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