Anti-Thymine Dimer antibody [H3] (ab10347)
- Datasheet
- References (7)
- Protocols
Overview
-
Product nameAnti-Thymine Dimer antibody [H3]
-
DescriptionMouse monoclonal [H3] to Thymine Dimer
-
Host speciesMouse
-
Tested applicationsSuitable 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
-
FormLiquid
-
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading... -
PurityProtein G purified
-
Primary antibody notesNon-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.
-
ClonalityMonoclonal
-
Clone numberH3
-
IsotypeIgG1
-
Research areas
Associated products
-
Compatible Secondaries
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
-
![Immunocytochemistry/ Immunofluorescence - Anti-Thymine Dimer antibody [H3] (ab10347)](https://www.abcam.com/ps/products/10/ab10347/Images/Thymine-Dimer-Primary-antibodies-ab10347-7.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Thymine Dimer antibody [H3] (ab10347)This image is courtesy of an anonymous Abreview.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.
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) »
