Overview

  • Product name
    Anti-Thymine Dimer antibody [H3]
  • Description
    Mouse monoclonal [H3] to Thymine Dimer
  • Specificity
    Monoclonal Anti-Thymine Dimer reacts specifically with the (5’-6’) cyclobutane type of homothymine or thymine-cytosine heterodimers. The antibody reacts with thymine dimers in single-stranded DNA, and has a lower affinity for the dimer in short oligonucleotides (a tail of minimum 10-20 thymine residues is required for efficient labeling of oligonucleotide probes).
  • Tested applications
    Suitable for: Southern Blot, ICC, Competitive ELISA, ELISA, ICC/IFmore details
  • Immunogen

    Chemical/ Small Molecule.

  • Positive control
    • ICC/IF: HeLa cells.
  • General notes


    Non-radioactive labeling of DNA is typically based on the enzymatic incorporation of modified nucleotides, carrying a small chemical moiety such as biotin, digoxigenin or fluorescein. These tags are subsequently detected by specific reagents such as streptavidin or a specific antibody coupled to a signal-producing enzyme. Although very efficient and reliable, labeling by in vitro polymerization is time-consuming, expensive, and may require various post-label purification steps to remove an excess of unincorporated precursors. An alternative strategy for DNA labeling, is based on the UV-induced formation of cyclobutane thymine dimers. Several methods have been described for the detection of thymine dimers, which are based on chromato-graphic analysis, and on biochemical analysis with endonucleases specific for UV-irradiated DNA. In addition, methods utilizing antibodies specific for pyrimidine dimers and other UV-induced DNA lesions have evolved, which permit the study of the induction and repair of these lesions without the requirement of in vivo radiolabeling of DNA. Photoimmunodetection, is a rapid, reliable and low-cost supplement to existing methods for nonradioactive DNA labeling. It enables a sensitive and non-radioactive method for labeling, detection, and quantification of high molecular weight (HMW) DNA fragments. The method is based on the introduction of thymine dimers into DNA after separa-tion by pulse field gel electrophoresis (PFGE), followed by detection with thymine dimer specific antibodies. The method does not require any enzymatic or chemical manipulation of the DNA sample. Monoclonal anti-bodies reacting specifically with thymine dimer, facilitate investigations on the apoptotic process and the role of UV-induced pyrimidine dimers in the process of photocarcinogenesis.

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: 15mM Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Protein G purified
  • Primary antibody notes
    Non-radioactive labeling of DNA is typically based on the enzymatic incorporation of modified nucleotides, carrying a small chemical moiety such as biotin, digoxigenin or fluorescein. These tags are subsequently detected by specific reagents such as streptavidin or a specific antibody coupled to a signal-producing enzyme. Although very efficient and reliable, labeling by in vitro polymerization is time-consuming, expensive, and may require various post-label purification steps to remove an excess of unincorporated precursors. An alternative strategy for DNA labeling, is based on the UV-induced formation of cyclobutane thymine dimers. Several methods have been described for the detection of thymine dimers, which are based on chromato-graphic analysis, and on biochemical analysis with endonucleases specific for UV-irradiated DNA. In addition, methods utilizing antibodies specific for pyrimidine dimers and other UV-induced DNA lesions have evolved, which permit the study of the induction and repair of these lesions without the requirement of in vivo radiolabeling of DNA. Photoimmunodetection, is a rapid, reliable and low-cost supplement to existing methods for nonradioactive DNA labeling. It enables a sensitive and non-radioactive method for labeling, detection, and quantification of high molecular weight (HMW) DNA fragments. The method is based on the introduction of thymine dimers into DNA after separa-tion by pulse field gel electrophoresis (PFGE), followed by detection with thymine dimer specific antibodies. The method does not require any enzymatic or chemical manipulation of the DNA sample. Monoclonal anti-bodies reacting specifically with thymine dimer, facilitate investigations on the apoptotic process and the role of UV-induced pyrimidine dimers in the process of photocarcinogenesis.
  • Clonality
    Monoclonal
  • Clone number
    H3
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab10347 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
Southern Blot Use a concentration of 0.5 - 1 µg/ml.
ICC Use at an assay dependent dilution.
Competitive ELISA Use at an assay dependent dilution.
ELISA Use at an assay dependent dilution.
ICC/IF Use at an assay dependent concentration.

Images

  • ab10347 staining Thymine Dimer in HeLa cells by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in formaldehyde, permabilized using 0.5% Triton X-100, blocked with 5% BSA for 15 minutes at 20°C, then incubated with ab10347 at a 1/250 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated rabbit anti mouse polyclonal, used at a 1/500 dilution.

    See Abreview

References

This product has been referenced in:
  • Torregrosa-Muñumer R  et al. Low doses of ultraviolet radiation and oxidative damage induce dramatic accumulation of mitochondrial DNA replication intermediates, fork regression, and replication initiation shift. Mol Biol Cell 26:4197-208 (2015). Read more (PubMed: 26399294) »
  • Deng Y  et al. A sunblock based on bioadhesive nanoparticles. Nat Mater 14:1278-85 (2015). Read more (PubMed: 26413985) »

See all 7 Publications for this product

Customer reviews and Q&As

Apologies, the expiration date should have been 30th September 2012. I have updated our records to reflect the correct date.

Please let me know if there is anything else I can help you with.

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (WI38 cell line)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
25 cells
Treatment
150 J/m2 UVC
Specification
WI38 cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 08 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human skin equivalent model, with and without UVB-)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 8.0 EDTA buffer (90¯C/40min)
Permeabilization
No
Specification
Human skin equivalent model, with and without UVB-
Blocking step
Biocare Medical Background SNIPER as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 24°C
Fixative
10% NBF
Username

Abcam user community

Verified customer

Submitted Jan 04 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (U2OS)
Specification
U2OS
Permeabilization
Yes - triton-X 100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 21 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Cell (u2os)
Specification
u2os
Type
Direct
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Andrew Cobb

Verified customer

Submitted Jul 16 2013

Thank you for your message.

I can confirm that Thymine Dimer is not a species specific target, and so this antibody should work in pig.

As far as we are aware, ab10347 has never been tested in IHC-P. All tested applications covere...

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Whole skin)
Specification
Whole skin
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
Yes - 0.1% Tween-20
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 11 2012

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

Having reviewed the protocol details, and after all the optimisation attempts, I am afraid ...

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I am sending you the new discount codes below, but would suggest if you encounter problems with using the codes for an online order, please give us a call so thatCustomer Service can help you with applying the codes to the order.

DISCOUNT COD...

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Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Acetone
Permeabilization
No
Username

Abcam user community

Verified customer

Submitted Jul 06 2012

1-10 of 19 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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