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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]


Monoclonal Antibodies against DNA Damage [PDF]

Go to "Antibodies against DNA Damage"

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.

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.

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
ELISA; 1:1500
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

Related Product

Cosmo Bio's new 6-4PP/ (6-4)Photoproducts ELISA Kit uses the highly sensitive and specific monoclonal anti-6-4PPs antibody clone 64M-2, established by Mori et al. 64M-2 is widely cited in the literature and is considered to be the gold standard antibody for 6-4PPs detection and quantification.Cosmo Bio's 6-4PPs ELISA Kit is the first and only commercially available ELISA kit using 64M-2. All kits components are optimized for high sensitivity 6-4PPs detection from cultured cells or skin epidermis.


Go To "High Sensitivity 6-4PP/ (6-4)Photoproducts ELISA kit"

Product List

Product Name Cat# Quantity Price

Anti 6-4PPs

CAC-NM-DND-002 1VIAL

¥ 44,000
$ 587
€ 440

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

CSR-NM-MA-K002 1KIT

¥ 100,000
$ 1334
€ 1000

Citation
  • Distinct Role of Sesn2 in Response to UVB-Induced DNA Damage and UVA-Induced Oxidative Stress in Melanocytes.
    Zhao B, Shah P, Qiang L, He TC, Budanov A, He YY.
    Photochem Photobiol. 2017 Jan;93(1):375-381. PMID: 27463837
  • A simple answer to complex questions: Caenorhabditis elegans as an experimental model for examining the DNA damage response and disease genes.
    Rieckher M, Bujarrabal A, Doll MA, Soltanmohammadi N, Schumacher B.
    J Cell Physiol. 2017 May 2. PMID: 28463453
  • Xeroderma pigmentosum group C protein interacts with histones: regulation by acetylated states of histone H3.
    Kakumu E, Nakanishi S, Shiratori HM, Kato A, Kobayashi W, Machida S, Yasuda T, Adachi N, Saito N, Ikura T, Kurumizaka H, Kimura H, Yokoi M, Sakai W, Sugasawa K.
    Genes Cells. 2017 Mar;22(3):310-327. PMID: 28233440
  • Identification of four novel XPC mutations in two xeroderma pigmentosum complementation group C patients and functional study of XPC Q320X mutant.
    Gu Y, Chang X, Dai S, Song Q, Zhao H, Lei P.
    Gene. 2017 Jun 29. pii: S0378-1119(17)30507-3. PMID: 28669926
  • Targeted Inactivation of DNA Photolyase Genes in Medaka Fish (Oryzias latipes).
    Ishikawa-Fujiwara T, Shiraishi E, Fujikawa Y, Mori T, Tsujimura T, Todo T.
    Photochem Photobiol. 2017 Jan;93(1):315-322. PMID: 27861979
  • Autophagy positively regulates DNA damage recognition by nucleotide excision repair.
    Qiang L, Zhao B, Shah P, Sample A, Yang S, He YY.
    Autophagy. 2016;12(2):357-68. PMID: 26565512
  • Oral Administration of Fermented Soymilk Products Protects the Skin of Hairless Mice against Ultraviolet Damage.
    Kano M, Kubota N, Masuoka N, Hori T, Miyazaki K, Ishikawa F.
    Nutrients. 2016 Aug 20;8(8). PMID: 27556484
  • Stabilization of Ultraviolet (UV)-stimulated Scaffold Protein A by Interaction with Ubiquitin-specific Peptidase 7 Is Essential for Transcription-coupled Nucleotide Excision Repair.
    Higa M, Zhang X, Tanaka K, Saijo M.
    J Biol Chem. 2016 Jun 24;291(26):13771-9. PMID: 27129218
  • Arabidopsis thaliana thymidine kinase 1a is ubiquitously expressed during development and contributes to confer tolerance to genotoxic stress.
    Pedroza-Garcia JA, Najera-Martinez M, de la Paz Sanchez M, Plasencia J.
    Plant Mol Biol. 2015 Feb;87(3):303-15. PMID: 25537647
  • SUMO and ubiquitin-dependent XPC exchange drives nucleotide excision repair.
    van Cuijk L, van Belle GJ, Turkyilmaz Y, Poulsen SL, Janssens RC, Theil AF, Sabatella M, Lans H, Mailand N, Houtsmuller AB, Vermeulen W, Marteijn JA.
    Nat Commun. 2015 Jul 7;6:7499. PMID: 26151477
  • Telomeres are partly shielded from ultraviolet-induced damage and proficient for nucleotide excision repair of photoproducts.
    Parikh D, Fouquerel E, Murphy CT, Wang H, Opresko PL.
    Nat Commun. 2015 Sep 9;6:8214. PMID: 26351258
  • SUMOylation of xeroderma pigmentosum group C protein regulates DNA damage recognition during nucleotide excision repair.
    Akita M, Tak YS, Shimura T, Matsumoto S, Okuda-Shimizu Y, Shimizu Y, Nishi R, Saitoh H, Iwai S, Mori T, Ikura T, Sakai W, Hanaoka F, Sugasawa K.
    Sci Rep. 2015 Jun 4;5:10984. PMID: 26042670
  • Direct exposure of non-equilibrium atmospheric pressure plasma confers simultaneous oxidative and ultraviolet modifications in biomolecules.
    Okazaki Y, Wang Y, Tanaka H, Mizuno M, Nakamura K, Kajiyama H, Kano H, Uchida K, Kikkawa F, Hori M, Toyokuni S.
    J Clin Biochem Nutr. 2014 Nov;55(3):207-15. PMID: 25411528
  • Chromatin retention of DNA damage sensors DDB2 and XPC through loss of p97 segregase causes genotoxicity.
    Puumalainen MR, Lessel D, Ruthemann P, Kaczmarek N, Bachmann K, Ramadan K, Naegeli H.
    Nat Commun. 2014 Apr 28;5:3695. PMID: 24770583

References
  • Simultaneous detection of ultraviolet B-induced DNA damage using capillary electrophoresis with laser-induced fluorescence.
    Guthrie JW, Limmer RT, Brooks EA, Wisnewski CC, Loggins-Davis ND, Bouzid A.
    Anal Chim Acta. 2015 Jan 1;853:676-81. PMID: 25467517
  • Perturbed gap-filling synthesis in nucleotide excision repair causes histone H2AX phosphorylation in human quiescent cells.
    Matsumoto M, Yaginuma K, Igarashi A, Imura M, Hasegawa M, Iwabuchi K, Date T, Mori T, Ishizaki K, Yamashita K, Inobe M, Matsunaga T.
    J Cell Sci. 2007 Mar 15;120(Pt 6):1104-12. PMID: 17327276
  • In vivo destabilization and functional defects of the xeroderma pigmentosum C protein caused by a pathogenic missense mutation.
    Yasuda G, Nishi R, Watanabe E, Mori T, Iwai S, Orioli D, Stefanini M, Hanaoka F, Sugasawa K.
    Mol Cell Biol. 2007 Oct;27(19):6606-14. PMID: 17682058
  • Neurons and astrocytes exhibit lower activities of global genome nucleotide excision repair than do fibroblasts.
    Yamamoto A, Nakamura Y, Kobayashi N, Iwamoto T, Yoshioka A, Kuniyasu H, Kishimoto T, Mori T.
    DNA Repair (Amst). 2007 May 1;6(5):649-57 PMID: 17236820
  • UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex.
    Sugasawa K, Okuda Y, Saijo M, Nishi R, Matsuda N, Chu G, Mori T, Iwai S, Tanaka K, Tanaka K, Hanaoka F.
    Cell. 2005 May 6;121(3):387-400. PMID: 15882621
  • Trichothiodystrophy fibroblasts are deficient in the repair of ultraviolet-induced cyclobutane pyrimidine dimers and (6-4)photoproducts.
    Nishiwaki Y, Kobayashi N, Imoto K, Iwamoto TA, Yamamoto A, Katsumi S, Shirai T, Sugiura S, Nakamura Y, Sarasin A, Miyagawa S, Mori T.
    J Invest Dermatol. 2004 Feb;122(2):526-32. PMID: 15009740
  • The total amount of DNA damage determines ultraviolet-radiation-induced cytotoxicity after uniformor localized irradiation of human cells.
    Imoto K, Kobayashi N, Katsumi S, Nishiwaki Y, Iwamoto TA, Yamamoto A, Yamashina Y, Shirai T, Miyagawa S, Dohi Y, Sugiura S, Mori T.
    J Invest Dermatol. 2002 Nov;119(5):1177-82. PMID: 12445209
  • DDB accumulates at DNA damage sites immediately after UV irradiation and directly stimulates nucleotide excision repair.
    Wakasugi M, Kawashima A, Morioka H, Linn S, Sancar A, Mori T, Nikaido O, Matsunaga T.
    J Biol Chem. 2002 Jan 18;277(3):1637-40. Epub 2001 Nov 8. PMID: 11705987
  • Quantitation and visualization of ultraviolet-induced DNA damage using specific antibodies: application to pigment cell biology.
    Kobayashi N, Katsumi S, Imoto K, Nakagawa A, Miyagawa S, Furumura M, Mori T.
    Pigment Cell Res. 2001 Apr;14(2):94-102. PMID: 11310797
  • Respective roles of cyclobutane pyrimidine dimers, (6-4)photoproducts, and minor photoproducts in ultraviolet mutagenesis of repair-deficient xeroderma pigmentosum A cells.
    Otoshi E, Yagi T, Mori T, Matsunaga T, Nikaido O, Kim ST, Hitomi K, Ikenaga M, Todo T.
    Cancer Res. 2000 Mar 15;60(6):1729-35. PMID: 10749146
  • Supranuclear melanin caps reduce ultraviolet induced DNA photoproducts in human epidermis.
    Kobayashi N, Nakagawa A, Muramatsu T, Yamashina Y, Shirai T, Hashimoto MW, Ishigaki Y, Ohnishi T, Mori T.
    J Invest Dermatol. 1998 May;110(5):806-10. PMID: 9579550
  • Three-dimensional visualization of ultraviolet-induced DNA damage and its repair in human cell nuclei.
    Nakagawa A, Kobayashi N, Muramatsu T, Yamashina Y, Shirai T, Hashimoto MW, Ikenaga M, Mori T.
    J Invest Dermatol. 1998 Feb;110(2):143-8. PMID: 9457909
  • Antigen structural requirements for recognition by a cyclobutane thymine dimer-specific monoclonal antibody.
    Komatsu Y, Tsujino T, Suzuki T, Nikaido O, Ohtsuka E.
    Nucleic Acids Res. 1997 Oct 1;25(19):3889-94. PMID: 9380513
  • High incidence of ultraviolet-B-or chemical-carcinogen-induced skin tumours in mice lacking the xeroderma pigmentosum group A gene.
    Nakane H, Takeuchi S, Yuba S, Saijo M, Nakatsu Y, Murai H, Nakatsuru Y, Ishikawa T, Hirota S, Kitamura Y, et al.
    Nature. 1995 Sep 14;377(6545):165-8. PMID: 7675085
  • A new photoreactivating enzyme that specifically repairs ultraviolet light-induced (6-4)photoproducts.
    Todo T, Takemori H, Ryo H, Ihara M, Matsunaga T, Nikaido O, Sato K, Nomura T.
    Nature. 1993 Jan 28;361(6410):371-4. PMID: 8426655
  • Melanin reduces ultraviolet-induced DNA damage formation and killing rate in cultured human melanoma cells.
    Kobayashi N, Muramatsu T, Yamashina Y, Shirai T, Ohnishi T, Mori T.
    J Invest Dermatol. 1993 Nov;101(5):685-9. PMID: 8228329
  • DNA damage in UV-irradiated human skin in vivo: automated direct measurement by image analysis (thymine dimers) compared with indirect measurement (unscheduled DNA synthesis) and protection by 5-methoxypsoralen.
    Potten CS, Chadwick CA, Cohen AJ, Nikaido O, Matsunaga T, Schipper NW, Young AR.
    Int J Radiat Biol. 1993 Mar;63(3):313-24. PMID: 8095281
  • Wavelength dependent formation of thymine dimers and (6-4) photoproducts in DNA by monochromatic ultraviolet light ranging from 150 to 365 nm.
    Matsunaga T, Hieda K, Nikaido O.
    Photochem Photobiol. 1991 Sep;54(3):403-10. PMID: 1784641
  • Simultaneous establishment of monoclonal antibodies specific for either cyclobutane pyrimidine dimer or (6-4)photoproduct from the same mouse immunized with ultraviolet-irradiated DNA.
    Mori T, Nakane M, Hattori T, Matsunaga T, Ihara M, Nikaido O.
    Photochem Photobiol. 1991 Aug;54(2):225-32. PMID: 1780359

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