DNA Methyltransferase 1 (DNMT1, DNMT, AIM, CXXC-type Zinc Finger Protein 9, CXXC9, DNA (Cytosine-5-)-methyltransferase 1, DNA MTase HsaI, FLJ16293, MCMT, M.HsaI, MGC104992)

Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting and development (1,2). Three families of mammalian DNA methyltransferases have been identified: \nDNMT1, DNMT2 and DNMT3 (1,2). DNMT1 is constitu-tively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methyla-tion patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic \nstem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltrans-ferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor main-tenance DNA methylation. DNMT1, DNMT3A and DNMT3B together form a protein complex that interacts with histone deacetylases (HDAC1, HDAC2, Sin3A), transcriptional re- \npressor proteins (RB, TAZ-1) and heterochromatin proteins (HP1, SUV39H1), to maintain proper levels of DNA methyla-tion and facilitate gene silencing (3-8). Improper DNA methylation contributes to diseased states such as cancer (1,2). Hypermethylation of promoter CpG islands within tumor suppressor genes correlates with gene silencing and the development of cancer. In addition, hypomethylation of bulk genomic DNA correlates with and may contribute to the \nonset of cancer. DNMT1, DNMT3A and DNMT3B are over-expressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast and stomach carcinomas (9-12). \n\nApplications:\nSuitable for use in Western Blot and Immunofluorescence (IC). Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000 Incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% \n Tween-20 at 4 degrees C with gentle shaking, overnight. \nImmunofluorescence (IC): 1:100\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

D3931-614

Size

100ul

Applications

IF, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, NHP

Form

Supplied as a liquid in 10mM sodium HEPES pH 7.5, 150mM NaCl, 0.1mg/ml BSA, 0.02% sodium azide and 50% glycerol.

P Type

Mab

Purity

Purified

Isotype

IgG

References

1. Hermann, A. et al. (2004) Cell. Mol. Life Sci. 61, 2571-2587, 2. Turek-Plewa, J. and Jagodzinski, P.P. (2005) Cell. Mol. Biol. Lett. 10, 631-647, 3. Kim, G.D. et al. (2002) EMBO J. 21, 4183-4195, 4. Fuks, F. et al. (2001) EMBO J. 20, 2536-2544, 5. Geiman, T.M. et al. (2004) Biochem. Biophys. Res. Commun. 318, 544-555, 6. Rountree, M.R. et al. (2000) Nat. Genet. 25, 269-277, 7. Pradhan, S. and Kim, G.D. (2002) EMBO J. 21, 779-788, 8. Fuks, F. et al. (2003) Nucleic Acids Res. 31, 2305-2312, 9. Mizuno, S. et al. (2001) Blood 97, 1172-1179, 10. Robertson, K.D. et al. (1999) Nucleic Acids Res. 27, 2291-2298, 11. Xie, S. et al. (1999) Gene 236, 87-95, 12. Kanai, Y. et al. (2001) Int. J. Cancer 91, 205-212.

Additional Info

Recongnizes endogenous levels of human total DNMT1 protein. Species crossreactivity: mouse, rat, monkey. Sequence Homology: bovine.