Anti-Topoisomerase I antibody (ab85038)

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Overview

  • Product name

    Anti-Topoisomerase I antibody
    See all Topoisomerase I primary antibodies

  • Description

    Rabbit polyclonal to Topoisomerase I

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Hamster, Cow, Monkey

  • Immunogen

    Synthetic peptide corresponding to Human Topoisomerase I aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab94391)

  • Positive control

    • This antibody gave a positive signal in the following human whole cell lysates: HeLa; Jurkat; HepG2; HEK293; MCF7; Caco2; SHSY5Y; NIH3T3; PC12.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab85038 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
IHC-P Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.

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

  • Western blot - Anti-Topoisomerase I antibody (ab85038)
    Western blot - Anti-Topoisomerase I antibody (ab85038)
    All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 90 kDa
    Observed band size: 90 kDa


    Exposure time: 30 seconds


    Human DNA topoisomerase 1 contains a potential sumoylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • Western blot - Anti-Topoisomerase I antibody (ab85038)
    Western blot - Anti-Topoisomerase I antibody (ab85038)
    All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 90 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 50 kDa, 55 kDa, 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • Immunoprecipitation - Anti-Topoisomerase I antibody (ab85038)
    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 - Anti-Topoisomerase I antibody (ab85038)
    Immunocytochemistry/ Immunofluorescence - Anti-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

Publishing research using ab85038? Please let us know so that we can cite the reference in this datasheet.

ab85038 has been referenced in 5 publications.

  • Lv C  et al. Analysis of topoisomerase I expression and identification of predictive markers for efficacy of topotecan chemotherapy in small cell lung cancer. Thorac Cancer 9:1166-1173 (2018). PubMed: 30058109
  • Bansal K  et al. The transcriptional regulator Aire binds to and activates super-enhancers. Nat Immunol 18:263-273 (2017). PubMed: 28135252
  • Guzman-Ayala M  et al. Chd1 is essential for the high transcriptional output and rapid growth of the mouse epiblast. Development 142:118-27 (2015). WB . PubMed: 25480920
  • Boichuk S  et al. Unbiased compound screening identifies unexpected drug sensitivities and novel treatment options for gastrointestinal stromal tumors. Cancer Res 74:1200-13 (2014). PubMed: 24385214
  • Qin H  et al. Transcriptional analysis of pluripotency reveals the Hippo pathway as a barrier to reprogramming. Hum Mol Genet 21:2054-67 (2012). WB . PubMed: 22286172

Customer reviews and Q&As

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Immunoprecipitation abreview for Anti-Topoisomerase I antibody

 Excellent
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (B-cell lymphocyte whole cell lysate)
Total protein in input
300 µg
Immuno-precipitation step
Protein A
Specification
B-cell lymphocyte whole cell lysate
Read More

Abcam user community

Verified customer

Submitted Mar 15 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Topoisomerase I antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibroblasts)
Permeabilization
Yes - 0.3% Triton X-100 in blocking buffer
Specification
fibroblasts
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

Submitted Nov 20 2017

Western blot abreview for Anti-Topoisomerase I antibody

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

Abcam user community

Verified customer

Submitted Feb 01 2017

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