Antibody to DNA methyltransferases (Dnmt Family)

Introduction

In mammals, epigenetic information is established and maintained via the postreplicative methylation of cytosine residues by the DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b.

Dnmt1 is required for maintenance methylation whereas Dnmt3a and Dnmt3b are responsible for de novo methylation. A systematic analysis of interactions among the Dnmt family members has revealed that DNMT3L interacts with the C-terminal domain of Dnmt3a and Dnmt3b. The interaction between Dnmt3-like (DNMT3L) and the C-terminal domains of Dnmt3a and Dnmt3b suggests a mechanism whereby the enzymatically inactive DNMT3L brings about the methylation of its substrate by recruiting an active methylase. Quantitative analysis of genomic cytosine methylation levels demonstrated a robust Dnmt activity for the de novo methyltransferases Dnmt3a and Dnmt3b. In addition, a weak but significant activity for Dnmt2 has been detected. Subsequent methylation tract analysis by genomic bisulphite sequencing revealed that Dnmt3 enzymes preferentially methylated CpG dinucleotides in a processive manner, whereas Dnmt2 methylated isolated cytosine residues in a non-CpG dinucleotide context.

Inappropriate over-expression of Dnmt can lead to inappropriate methylation of DNA and thus, mis¬regulation of the cell's genes. If, for example, inappropriate methylation occurs on tumour suppressor genes, expression of the tumour suppressor genes is turned off, and cancer may develop. The methylation of tumour suppressor genes has been observed to occur in a wide variety of human cancers.

Antibody Panel to the Dnmt Family

Acris Antibodies offers a range of monoclonal and polyclonal antibodies to Dnmt1, Dnmt2, Dnmt3a/3b and Dnmt3-like (DNMT3L). All antibodies are suitable for Western blot, ELISA and / or Immunoprecipitation/ Immunohistochemical.

Fig. 1: Western blot of Dnmt1 on HeLa cell Nuclear extract using antibody SP5327P. This antibody detects an ~180 kDa protein. A doublet between 40 and 45 kDa is also seen .

Fig. 2: Western blot analysis for Dnmt1 using SM7026P at 2 µg/ml against 10 µg of cell lysates of mouse ES cell.

Fig. 3: Western blot analysis for Dnmt3b using SM7029P at 2 µg/ml against 10 µg of cell lysates of 293 transfected with mouse Dnmt3a (lane 1) and Dnmt3b (lane 2). A protein band of ~110 kDa is only detected in 293 cells transfected with mouse Dnmt3b cDNA.

Fig. 4: IHC analysis of Dnmt1 using SM7026P at 2 µg/ml on formalin-fixed paraffin-embedded medullar kidney tissue sections.

Fig. 5: Immuno-staining of Dnmt3a using antibody SM7028P at 5 ug/ml dilution on cultured HeLa cells. The nucleic staining is shown in most of the cells.

Table of Antibody Panel to Dnmt Family

DNA methyltransferases (Dnmt1, Dnmt2, Dnmt3a/3b), Dnmt-like (DNMT3L) antibody.

Literature

Cheng X, Blumenthal RM Mammalian DNA methyltransferases: a structural perspective. Structure. 2008 Mar;16(3):341-50.

Mund C, Musch T, Strödicke M, Assmann B, Li E, and Lyko F. Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases. Biochem J. 2004 Mar 15; 378 (Pt3): 763-8.

Margot JB, Ehrenhofer-Murray AE, Leonhardt H. Interactions within the mammalian methyltransferase family. BMC Mol Biol. 2003 May 30; 4:7

Liu K, Wang YF, Cantemir C, Muller MT. Endogenous assays of DNA methyltransferases: Evidence for differential activities of DNMT1, DNMT2, and DNMT3 in mammalian cells in vivo. Mol Cell Biol. 2003 Apr; 23(8): 2709-19.

Kareta MS, Botello ZM, Ennis JJ, Chou C, Chédin F. Reconstitution and mechanism of the stimulation of de novo methylation by human DNMT3L. J Biol Chem. 2006 Sep 8;281(36):25893-902.

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