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Antibodies
UV-Induced DNA Damage
(monoclonals)

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Antibodies: DNA Replication, Recombination, Repair
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see also: CPD and 6-4 PP ELISA Kits (featuring mAb clones
TDM-2 and 64M-2!)
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Anti Cyclobutane Pyrimidine Dimers (CPDs), clone TDM-2
Anti (6-4) Photoproducts (6-4 PPs), clone 64M-2
Anti Dewar Photoproducts (Dewar PPs), clone DEM-1

  • Quantitate photoproducts by ELISA or visualize by immunofluorescence.
  • Specific for nominal UV-induced DNA lesion.
  • Detect photoproducts in single stranded DNA or oligos (8bp or greater).
  • Reacts with photoproducts from any species.
  • Widely cited.
Ordering Information
Name CBU Price Alternate Name Host Clone Applications Size Cat No
Anti CPDs $587 Anti Cyclobutane Pyrimidine Dimers MS TDM-2 ELISA, IC 100 ul CAC-NM-DND-001
Anti 6-4PPs $587 Anti 6-4 Photoproducts MS 64M-2 ELISA, IC 100 ul CAC-NM-DND-002
Anti DewarPPs $587 Anti Dewar photoproducts MS DEM-1 ELISA, IC 100 ul CAC-NM-DND-003
Background

Prolonged exposure to solar UV radiation may result in acute and chronic health effects on the skin, eye, and immune system, including skin cancers. These harmful effects are suggested to deeply relate to DNA damage.

The major types of DNA damage induced by solar UV radiation are cyclobutane pyrimidine dimers (CPDs), (6–4) photoproducts (6-4PPs), and Dewar photoproducts (DewarPPs), which are formed between adjacent pyrimidine nucleotides on the same strand of DNA. These helix-distorting DNA lesions are repaired exclusively by a nucleotide excision repair system in humans.

Mori et. al. have developed and characterized monoclonal antibodies specific for CPDs or 6-4PPs (1) while Matsunaga et. al. have established and characterized monoclonal antibodies against DewarPPs (2). These antibodies enable quantitation of photoproducts in DNA purified from cultured cells or from skin epidermis using an enzyme-linked immunosorbent assay (ELISA), and to visualize and measure photoproducts in DNA of cultured cells or skin by indirect immunofluorescence. These capabilities will contribute to understanding the molecular mechanism of cellular responses to UV and DNA damage in many research fields including cancer research, photobiology, dermatology, ophthalmology, immunology, and cosmetology.

(1) Toshio Mori, Misa Nakane, Tsuyoshi Hattori, Tsukasa Matsunaga, Makoto Ihara, Osamu Nikaido. Simultaneous establishment of monoclonal antibodies specific for either cyclobutane pyrimidine dimer or (6-4) photoproduct from the same mouse immunized with ultraviolet-irradiated DNA. Photochem. Photobiol., 54: 225-232 (1991). PMID 1780359

(2) Tsukasa Matsunaga, Yuri Hatakeyama, Michi Ohta, Toshio Mori and Osamu Nikaido. Establishment and characterization of a monoclonal antibody recognizing the Dewar isomers of (6-4) photoproducts. Photochem. Photobiol., 57: 934-940 (1993). PMID 8367534

Applications
Immunohistochemistry  
 
The technique of micropore UV irradiation combined with fluorescent antibody labeling is very powerful for examining whether a protein of interest is recruited to the sites of UV-induced DNA damage. Micropore UV irradiation induces UV-damage at localized areas of nuclei using a polycarbonate isopore membrane filter. The polycarbonate blocks UV radiation, and cells are exposed only through the 5 µm pores of the filter. 0.5 h after micropore UV irradiation, cells were fixed and Immunofluorescent double staining for DNA damage and repair protein were performed.  
 
Cells were doubly stained for XPB and for CPD 0.5 h after local UV irradiation. In normal MSU-1 cells, XPB foci overlapped with the corresponding CPD foci, indicating that XPB is quickly recruited to the sites of DNA damage for repair. In contrast, no or less bright XPB foci at the DNA damage sites were observed in repair deficient TTD cell lines.  
ELISA  
 
Genomic DNA is purified from UV-damaged cells and denatured DNA is coated on wells of 96-well plate. The binding of TDM-2 or 64M-2 to DNA damage is detected by sequential treatment with biotinylated 2nd antibody and streptavidin-peroxidase. Then, the absorbance of colored products derived from OPD is measured at 492 nm.  
 
Normal human cells repair 90% of the initial 6-4PP within 3 h after UV irradiation, while they remove 50% of the initial CPD within 24 h. Both damage are repaired by the same nucleotide excision repair (NER) pathway, but 6-4PP forms bigger distortion in DNA than CPD does, resulting in much more efficient repair. In contrast, repair deficient XP-C cells can not repair both damage at all.  
 
Related Products      
Product Name (click for more info) Cosmo Bio USA Price Unit Cat. No.
(click for datasheet)
High Sensitivity CPD ELISA Kit featuring mAb TDM-2 $1,104 1kit (96 tests) CSR-NM-MA-K001
UVC-irradiated DNA samples $331 1 set CSR-NM-MA-R010
Protamine Sulfate Coated ELISA Plate $23 1 plate CSR-NM-MA-P001
Protamine Sulfate Coated ELISA Plate $106 5 plates CSR-NM-MA-P002
Protamine Sulfate Coated ELISA Plate $199 10 plates CSR-NM-MA-P003
8-OHdG Check ELISA kit $1,130 96 WELL NNS-KOG-200SE-EX
Highly Sensitive 8-OHdG Check ELISA kit $1,130 96 WELL NNS-KOG-HS10E-EX
       
Antibodies to DNA Replication, Recombination, Damage and Repair Proteins      

 

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