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DNA Damage Antibody for research on DNA Damage and Repair

Anti (6-4) photoproducts (6-4 PPs) monoclonal antibody [Clone : 64M-2]


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Background

DNA damage in cells exposed to ultraviolet (UV) radiation plays significant roles in cell-cycle arrest, activation of DNA repair, cell killing, mutation, and neoplastic transformation.

The major types of DNA damage induced by UVB (280-315 nm, component of sunlight) and by UVC (200-280nm) are cyclobutane pyrimidine dimers (CPDs) and (6-4) photoproducts (6-4PPs), which are formed between adjacent pyrimidine nucleotides on the same strand of DNA.

Approximately 70-80% of UV-induced DNA damage is CPDs and the remaining is 6-4PPs and Dewar isomer of 6-4PPs. These types of DNA lesions are repaired by nucleotide excision repair (NER) system in normal human cells. Mori et al (19) have established monoclonal antibodies specific for CPDs or 6-4PPs.

These antibodies enable one to quantitate photoproducts in DNA purified from cultured cells or from the skin epidermis using an enzyme-linked immunosorbent assay (ELISA) and to visualize and measure photoproducts in DNA in cultured cells or the skin using indirect immunofluorescence (IIF). This technology would contribute to understanding of molecular mechanisms of cellular responses to UV and DNA damage in many research fields including cancer research, photobiology, dermatology, ophthalmology, immunology, and cosmetology.

Monoclonal Antibodies against DNA DamagePDF

[SOURCE]
This hybridoma was established by fusion of mouse myeloma cells with Balb/c mouse splenocytes immunized with methylated BSA conjugated with calf thymus DNA which was irradiated with UVC. This hybridoma (clone 64M-2) culture supernatant was collected and precipitated with ice-cold ammonium sulfate. After centrifugation, the pellet dissolved in small volume of double-distilled water was dialysed against PBS. The dialysate was then lyophilized.

[REACTIVITY]
1) The antibodies bind to 6-4PPs in single-stranded DNA.
2) The antibodies bind to 6-4PPs formed in every dipyrimidine sequence (TT, TC, CT and CC).
3) The antibodies stably bind to 6-4PPs formed in oligonucleotides consisting of more than eight bases.

[SPECIES CROSS REACTIVITY]
The antibodies can bind to 6-4PPs in denatured DNA from all organisms from bacteria to human.

 

Features

React specifically with damaged DNA
Research applications include ELISA and IC
Useful for research into DNA damage repair mechanisms
Visualizes the DNA damage repair process
Applicable to a broad range of research interests:photobiology, optical medicine, cancer, etc.

Application

Immunocytochemistry; 1:300 PROTOCOLS<ELISA>PDF
ELISA; 1:1500@PROTOCOLS< Immunofluorescence microscopy >PDF
Western blotting; Not tested
Immunoprecipitation; Not tested
Immunohistochemistry; Not tested
Flow cytometry; Not tested

Induction and repair of UV-induced DNA damage in normal human cells measured by ELISA

Visualization of UV-induced DNA damage by using immunofluorescence

Visualization of UV-induced 6-4PP and its repair

<Reference>
1. Yamamoto, A., et al., DNA Repair, 6, 649-657 (2007).
2. Matsumoto, M.,et al., J. Cell Sci., 120, 1104-1112 (2007).
3. Yasuda, G., et al., Mol. Cell. Biol., 27, 6606-6614 (2007).
4. Sugasawa, K., et al., Cell 121, 387-400 (2005).
5. Nishiwaki, Y., et al., J. Invest. Dermatol. 122, 526-532 (2004)
6. Imoto, K., et al., J. Invest. Dermatol. 119, 1177-7782 (2002)
7. Wakasugi, M., et al., J. Biol. Chem., 277, 1637-1640 (2002).
8. Kobayashi, N., et al., Pigment Cell Res. 14, 94-102 (2001)
9. Katsumi, S., et al., J. Invest. Dermatol. 117, 1156-1161 (2001)
10. Otoshi, E., et al., Cancer Res. 60, 1729-1735 (2000)
11. Nakagawa, A., et al., J. Invest. Dermatol. 110, 143-148 (1998)
12. Kobayashi, N., et al., J. Invest. Dermatol. 110, 806-810 (1998)
13. Komatsu, Y., et al., Nucleic Acids Res. 25, 3889-3894 (1997)
14. Nakane, H., et al., Nature 377, 165-168 (1995)
15. Todo, T., et al., Nature 361, 371-374 (1993)
16. Kobayashi, N., et al., J. Invest. Dermatol. 101, 685-689 (1993)
17. Potten, C.S., et al., Int. J. Radiat. Biol. 63, 313-324 (1993)
18. Matsunaga, T., et al., Photochem. Photobiol. 54, 403-410 (1991)
19. Mori, T., et al., Photochem. Photobiol. 54, 225-232 (1991)

Product List

Host : MS, Clone : 64M-2, Application : ELISA/IC


Product Name Cat# Quantity Price

Anti 6-4PPs (Mouse)  DataSheet

CAC-NM-DND-002

1 VIAL ¥44,000
$587

High Sensitivity 6-4PP/ (6-4) photoproduct ELISA kit DataSheet ,

CSR-NM-MA-K002

1 KIT ¥100,000
$1334
Anti CPDs, CAC-NM-DND-001, Host : MS, Clone : TDM-2, ELISA/IC
Anti Dewar PPs, CAC-NM-DND-003, Host : MSClone : DEM-1, ELISA/IC

Product Name Cat# Quantity Price

Anti CPDs (Mouse)  DataSheet

CAC-NM-DND-001

1 VIAL ¥44,000
$587

Anti DewarPPs, Human (Mouse)  DataSheet

CAC-NM-DND-003

1 VIAL ¥44,000
$587

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.



“Anti (6-4) photoproducts (6-4 PPs) monoclonal antibody [Clone : 64M-2]” also belongs to the following categories.