Overview

  • Product name

    Anti-Topoisomerase I antibody [EPR5375]
    See all Topoisomerase I primary antibodies
  • Description

    Rabbit monoclonal [EPR5375] to Topoisomerase I
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Topoisomerase I aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB; Jurkat, SW480, MCF7, HepG2, Neuro-2a, PC-12, and K562 cell lysates; IHC-P: Human breast carcinoma, Human colonic carcinoma tissues, Human clear cell carcinoma of kidney and Mouse kidney, and Rat colon tissue; ICC/IF: MCF7 cells;
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109374 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
WB 1/10000 - 1/50000. Predicted molecular weight: 91 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/500.

For unpurified use at 1/100 - 1/250

  • Application notes
    Is unsuitable for IP.
  • Target

    • 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
      modifications

      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.
    • 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
    • Database links

    • 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
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling Topoisomerase I with Purified ab109374 at 1:100 dilution (1.29 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Topoisomerase I with Purified ab109374 at 1/500 dilution (0.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Topoisomerase I with Purified ab109374 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 80% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling Topoisomerase I with Purified ab109374 at 1:100 dilution (1.29 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell carcinoma of kidney tissue sections labeling Topoisomerase I with Purified ab109374 at 1:100 dilution (1.29 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Overlay histogram showing HepG2 cells stained with ab109374 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109374, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    • All lanes : Anti-Topoisomerase I antibody [EPR5375] (ab109374) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : SW480 cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 91 kDa

    • Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue using ab109374 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded Human colonic carcinoma tissue using ab109374 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunofluorescent staining of MCF7 cells using ab109374 at a dilution of 1/100.

    References

    This product has been referenced in:

    • Zyner KG  et al. Genetic interactions of G-quadruplexes in humans. Elife 8:N/A (2019). Read more (PubMed: 31287417) »
    • Aumailley L  et al. Vitamin C alters the amount of specific endoplasmic reticulum associated proteins involved in lipid metabolism in the liver of mice synthesizing a nonfunctional Werner syndrome (Wrn) mutant protein. PLoS One 13:e0193170 (2018). Read more (PubMed: 29494634) »
    See all 18 Publications for this product

    Customer reviews and Q&As

    1-9 of 9 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Blood)
    Gel Running Conditions
    Non-reduced Denaturing (8% GEL)
    Loading amount
    30 µg
    Treatment
    5 uM KML001 for 48h
    Specification
    Blood
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Mr. Dechao Cao

    Verified customer

    Submitted Jun 11 2019

    Application
    Other
    Sample
    Human Cell lysate - other (top1 cleavage complex purify by CscL gradient)

    Ms. Cristina Bernal

    Verified customer

    Submitted Apr 01 2019

    This product is known to not work in this application or species.
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Total protein in input
    500 µg
    Immuno-precipitation step
    Protein A/G
    Specification
    U2OS cells

    Abcam user community

    Verified customer

    Submitted May 12 2017

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEF cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    20 µg
    Specification
    MEF cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Mar 28 2017

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    50 µg
    Specification
    U2OS cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 05 2016

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (fibroblast)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    40 µg
    Specification
    fibroblast
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Aug 05 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (fibroblast)
    Permeabilization
    Yes - block buffer contains 0.3% TritonX-100, 1H, RT
    Specification
    fibroblast
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 29 2016

    Answer

    Merci de nous avoir contactés.

    La concentration de l'anti-Topoisomerase I ab109374, lot numéro GR49853-3 est 0.109 mg/mL.

    J'espère que cette information vous sera utile. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Read More

    Answer

    Thank you for your enquiry. Please find enclosed the two requested CoC for ab3380 (Lot NO.:GR56451-1) and ab109374 (Lot NO.:GR49853-2) as attachments. I hope this helps and if I can assist further, please do not hesitate to contact me.  

    Read More

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