Cayman Europe | DNA Methylation EIA Kit | Description • 5-methyl-2-deoxy Cytidine ELISA Kit

Limit of detection: 80% B/B₀: ~3 ng/ml · Sensitivity: 50% B/B₀: ~12 ng/ml · DNA methylation is an important epigenetic process regulating gene expression. Methylation occurs on carbon 5 of 2-deoxy cytidine yielding the modified base 5-methyl-2-deoxy cytidine. The methylation pattern of cells is tightly regulated during development with the methylation profile being transmitted from parent to daughter cells during cell division.¹ Methylation results in long-term silencing of genes, while unmethylated regions of DNA can be actively transcribed.^1,2 One region of the genome of particular interest in regard to methylation is CpG islands. CpG islands are regions approximately 200 bp to several thousand bp in length which often span the promoter and first few exons of many housekeeping and tumor suppressor genes. These regions remain essentially unmethylated throughout development.¹ As the name implies, the CpG dinucleotide is overrepresented in CpG islands, with the frequency being approximately five times greater in CpG islands than in the remainder of the genome.¹ It is well established that alterations in DNA methylation are a common feature of cancer.³ In addition to global genomic hypomethylation, there are also discrete areas of dense hypermethylation particularly in the normally unmethylated CpG islands.³ Because many tumor suppressor genes contain CpG islands, these genes are among those silenced by hypermethylation. The first report of methylation of a tumor suppressor CpG island was Retinoblastoma (Rb) which was discovered in 1989.⁴ Interestingly, the pattern and level of gene hypermethylation for a given cancer is specific to that malignancy, with cancer of the GI tract tending to show a greater level of hypermethylation than do other cancers. In the digestive tract, methylation of genes, including the estrogen receptor, occurs as a normal part of the aging process.^5,6,7 During carcinogenesis, this age-related methylation may progress to hypermethylation.^5,6 Global changes in methylation can be quantified by measuring plasma or urinary levels of 5-methyl-2-deoxy cytidine.^8,9,10 These changes in methylation can provide valuable information about cancer status of an individual. For example, patients with leukemia excrete significantly elevated levels of 5-methyl-2-deoxy cytidine compared to healthy individuals.⁸ Global methylation within tissues can be measured in a similar manner, allowing study of tissue-specific changes that occur as a result of differentiation, aging, or carcinogenesis.¹¹ DNA methylation is regulated by a family of DNA methyltransferases (DNMTs), with DNMT1, DNMT3a, and DNMT3b all implicated in carcinogenesis.¹² Three cytosine nucleoside analogs (azacitidine, decitabine, and zebularine) which incorporate into DNA during synthesis are being actively investigated as anti-cancer drugs.¹³ DNMTs bind irreversibly to these cytidine analogs, resulting in suppression of methylation and the possibility that genes which had been inappropriately methylated may resume their normal function.¹³ Cayman’s 5-methyl-2-deoxy Cytidine EIA is a competitive assay that can be used for the quantification of 5-methyl-2-deoxy cytidine in urine, culture supernatants, plasma, and other sample matrices. The EIA typically displays IC₅₀ (50% B/B₀) and IC₈₀ (80% B/B₀) values of approximately 12 and 3 ng/ml, respectively.

1 Clark, S.J., Melki, J. DNA methylation and gene silencing in cancer: Which is the guilty party?. Oncogene 21 5380-5387 (2002).

2 Robertson, K.D. DNA, methylation and chromatin-unraveling the tangled web. Oncogene 21 5361-5379 (2002).

3 Ushijima, T. Detection and interpretation of altered methylation patterns in cancer cells. Cancer 5 223-231 (2005).

4 Greger, V., Passarge, E., Höpping, W., et al. Epigenetic changes may contribute to the formation and spontaneous regression of retinoblastoma. Hum Genet 83 155-158 (1989).

5 Ahuja, N., Li, Q., Mohan, A.L., et al. Aging and DNA methylation in colorectal mucosa and cancer. Cancer Res 58 5489-5494 (1998).

6 Issa, J.J., Ottaviano, Y.L., Celano, P., et al. Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon. Nature Genet 7 536-540 (1994).

7 Waki, T., Tamura, G., Sato, M., et al. Age-related methylation of tumor suppressor and tumor-related genes: An analysis of autopsy samples. Oncogene 22 4128-4133 (2003).

8 Itoh, K., Aida, S., Ishiwata, S., et al. Immunochemical detection of urinary 5-methyl-2'-deoxycytidine as a potential biologic marker for leukemia. Clin Chim Acta 234 37-45 (1995).

9 Mizugaki, M., Itoh, K., Yamaguchi, T., et al. Preparation of a monoclonal antibody specific for 5-methyl-2'-deoxycytidine and its application for the detection of DNA methylation levels in human peripheral blood cells. Biol Pharm Bull 12 1537-1540 (1996).

10 Reynaud, C., Bruno, C., Boullanger, P., et al. Monitoring of urinary excretion of modified nucleosides in cancer patients using a set of six monoclonal antibodies. Cancer Lett (1991).

11 Wilson, V.L., Smith, R.A., Autrup, H., et al. Genomic 5-methylcytosine determination by 32P-postlabeling analysis. Anal Biochem 152 275-284 (1986).

12 Linhart, H.G., Lin, H., Yamada, Y. DNMT3b promotes tumorigenesis in vivo by gene-specific de novo methylation and transcriptional silencing. Genes Dev 21 3110-3122 (2007).

13 Flotho, C., Claus, R., Batz, C., et al. The DNA methyltransferase inhibitors azacitidine, decitabine and zebularine exert differential effects on cancer gene expression in acute myeloid leukemia cells. Leukemia (2009).

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589324 DNA Methylation EIA Kit

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