RecombinantRabMAb

Recombinant Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5375] to Topoisomerase I - BSA and Azide free
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free
    See all Topoisomerase I primary antibodies

  • Description

    Rabbit monoclonal [EPR5375] to Topoisomerase I - BSA and Azide free

  • 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 corresponding to residues at the N-terminus of Human Topoisomerase I

  • Positive control

    • 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

    Ab219735 is the carrier-free version of ab109374. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab219735 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab219735 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 Use at an assay dependent concentration. Predicted molecular weight: 91 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
ICC/IF Use at an assay dependent concentration.
  • 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

      see all

    • 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) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

      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 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.
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374)

    • Flow Cytometry - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Flow Cytometry - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      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).
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374)
    • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

      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.
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

      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 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.
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

      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 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.
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374)

    • Flow Cytometry - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Flow Cytometry - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374).

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374).

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Immunofluorescent staining of MCF7 cells using ab109374 at a dilution of 1/100.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109374).

    • Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)
      Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free (ab219735)

    References (5)

    Publishing research using ab219735? Please let us know so that we can cite the reference in this datasheet.

    ab219735 has been referenced in 5 publications.

    • 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). WB . PubMed: 24787137
    • Mamouni K  et al. RhoB promotes ?H2AX dephosphorylation and DNA double-strand break repair. Mol Cell Biol 34:3144-55 (2014). PubMed: 24912678
    • Kouzine F  et al. Transcription-dependent dynamic supercoiling is a short-range genomic force. Nat Struct Mol Biol 20:396-403 (2013). ChIP ; Human . PubMed: 23416947
    • Eliasof S  et al. Correlating preclinical animal studies and human clinical trials of a multifunctional, polymeric nanoparticle. Proc Natl Acad Sci U S A 110:15127-32 (2013). WB ; Human . PubMed: 23980155
    • Chen CY  et al. Rescue of the genetically engineered Cul4b mutant mouse as a potential model for human X-linked mental retardation. Hum Mol Genet 21:4270-85 (2012). Mouse . PubMed: 22763239

    Customer reviews and Q&As

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    ChIP abreview for Anti-Topoisomerase I antibody [EPR5375] - BSA and Azide free

     Excellent
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (Colorectal carcinoma)
    Negative control
    IP using IgG antibody
    Specification
    Colorectal carcinoma
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 30 minute(s) and 0 second(s)
    Specification of the cross-linking agent: DSG
    Read More

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    Verified customer

    Submitted Nov 11 2019

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