Overview

  • Product name
  • Description
    Rabbit polyclonal to TDP1
  • Tested applications
    Suitable for: IP, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide derived from residues 50 - 150 of Human TDP1.

  • Positive control
    • This antibody gave a positive signal when tested against TDP1 recombinant protein and jurkat nuclear cell lysate.

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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab4166 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
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 21041670
WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 68 kDa).

Abcam recommends blocking with 1-3% Milk.

IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. 20 min in boiling Tris-EDTA solution, see Abreview by Frédéric Leduc.

Target

  • Function
    DNA repair enzyme that can remove a variety of covalent adducts from DNA through hydrolysis of a 3'-phosphodiester bond, giving rise to DNA with a free 3' phosphate. Catalyzes the hydrolysis of dead-end complexes between DNA and the topoisomerase I active site tyrosine residue. Hydrolyzes 3'-phosphoglycolates on protruding 3' ends on DNA double-strand breaks due to DNA damage by radiation and free radicals. Acts on blunt-ended double-strand DNA breaks and on single-stranded DNA. Has low 3'exonuclease activity and can remove a single nucleoside from the 3'end of DNA and RNA molecules with 3'hydroxyl groups. Has no exonuclease activity towards DNA or RNA with a 3'phosphate.
  • Tissue specificity
    Ubiquitously expressed. Similar expression throughout the central nervous system (whole brain, amygdala, caudate nucleus, cerebellum, cerebral cortex, frontal lobe, hippocampus, medulla oblongata, occipital lobe, putamen, substantia nigra, temporal lobe, thalamus, nucleus accumbens and spinal cord) and increased expression in testis and thymus.
  • Involvement in disease
    Spinocerebellar ataxia, autosomal recessive, with axonal neuropathy
  • Sequence similarities
    Belongs to the tyrosyl-DNA phosphodiesterase family.
  • Post-translational
    modifications
    Phosphorylated on serine and/or threonine residues, but not on tyrosine residues.
  • Cellular localization
    Nucleus. Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • AI838772 antibody
    • AW493413 antibody
    • FLJ11090 antibody
    • MGC104252 antibody
    • MGC112732 antibody
    • RP24-311F12.2 antibody
    • SCAN1 antibody
    • TDP1 antibody
    • TYDP antibody
    • TYDP1_HUMAN antibody
    • Tyr-DNA phosphodiesterase 1 antibody
    • Tyrosyl-DNA phosphodiesterase 1 antibody
    see all

Images

  • Anti-TDP1 antibody (ab4166) at 1 µg/ml (blocked with 3% milk) + Jurkat nuclear extract lysate (ab14844) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 68 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)


    Exposure time : 4 minutes
  • Lane 1 : Molecular weight markers
    Lanes 2 - 5 : Anti-TDP1 antibody (ab4166) at 1/1000 dilution

    Lane 1 : As above
    Lane 2 : 1 ng purified recombinant human TDP1
    Lane 3 : 3 ng purified recombinant human TDP1
    Lane 4 : 6 ng purified recombinant human TDP1
    Lane 5 : 6 micrograms total protein from MCF7 human cell extract

    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 68 kDa
    Observed band size : 68 kDa

    This image is courtesy of Keith Caldecott, University of Sussex

    Western blot using ab4166 at 1/1000 dilution for 1 hr at room temp. The lanes are as follows, from left to right. The blot was developed using ECL.

    1. markers
    2. 1ng purified recombinant human TDP1
    3. 3ng purified recombinant human TDP1
    4. 6ng purified recomninant human TDP1
    5. 6 micrograms total protein from MCF7 human cell extract.

    ab4166 was incubated for 1 hr at room temp.
  • ab4166 staining TDP1 in MCF-7 cells by Immunocytochemistry/ Immunofluorescence.
    Briefly, cells were incubated in medium containing 100 nM Mitotracker Red 580 (red) at 37 °C for 30 minutes and fixed with 4% paraformaldehyde. Primary antibody against TDP1 was detected with anti-rabbit IgG secondary antibodies labeled with Alexa 488 (green). Nuclei were stained with DAPI (blue).

    TDP1 localizes to the mitochondria. Immunofluorescence images showing colocalization of TDP1 (green) and mitochondria (red). (Upper) Cellular distribution of TDP1 at lower magnification. The boxed area is shown magnified below (Lower) to better show colocalization of TDP1 and mitochondria.

References

This product has been referenced in:
  • Meisenberg C  et al. Epigenetic changes in histone acetylation underpin resistance to the topoisomerase I inhibitor irinotecan. Nucleic Acids Res 45:1159-1176 (2017). WB ; Human . Read more (PubMed: 28180300) »
  • Chiang SC  et al. Mitochondrial protein-linked DNA breaks perturb mitochondrial gene transcription and trigger free radical-induced DNA damage. Sci Adv 3:e1602506 (2017). Read more (PubMed: 28508041) »

See all 19 Publications for this product

Customer reviews and Q&As

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 25 2017

Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Human Cell lysate - whole cell (Human primary fibroblasts)
Specification
Human primary fibroblasts
Total protein in input
30 µg
Username

Abcam user community

Verified customer

Submitted Jan 19 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: RT°C
Sample
Human Cell (HeLa cells)
Specification
HeLa cells
Permeabilization
Yes - 0.2% Triton-X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 24 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient Tris-Gly gel)
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Blocking step
LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 19 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient Tris-Gly gel)
Sample
Mouse Cell lysate - whole cell (MEFs)
Specification
MEFs
Blocking step
LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 19 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (testis)
Specification
testis
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 37°C
Username

Mr. Frédéric Leduc

Verified customer

Submitted Jan 27 2009

I was just passed your email about the TDP1 antibody ab4166. I am not sure what you were told before, but we have just published the antibody on the website this week (www.abcam.com/ab4166). So you can go ahead and buy it now. Our collaborator whom we ...

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