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code:10-201

DNA methylation is significant for epigenetic regulation of gene expression X chromosome inactivation genomic imprinting and development. Abberant methylation patterns are associated with certain human tumors and developmental abnormalities. In vertebrates there are two types of DNA methyltransferase activities; de novo and maintenance types. Two DNA methyltransferases Dnmt3a and Dnmt3b are responsible for the creation of methylation patterns at an early stage of embryogenesis (de novo-type) while Dnmt1 is responsible for the maintenance of methylation patterns during replication. Dnmt1 favors to methylate the hemimethylated DNA and preferentially methylates one strand of the double-stranded DNA during its processive methylation. This product mouse Dnmt1 deleting the N-terminal 290 amino acid residues was expressed using a baculovirus expression system and purified by Prof. S. Tajima and Dr. I. Suetake of Osaka University (ref.2). Since a trace of baculovirus contamination in the purified Dnmt1 the material and the container have to be sterilized at 121 C for 20 min before disposal according to Cartagena Protocol on Biodiversity.

Applications
1)In vitro metylation of cytosine residues in hemimethylated      DNA at 5’….CG…3’. (ref. 12)
2)Antigen for anti-mammalian Dnmt1 antibodies.

Form: 0.5mg protein/ml in 0.2M NaCl 10mM HEPES (pH 7.4) 50% glycerol
Definition of specific activity: 1 unit is defined as the amount of the enzyme that transfer 1 pmole of
methyl group to poly dI-dC substrate during 30 minutes at 37℃

Specific activity:17 units/ul

Quality Assurance: Greater than 95% protein determined by SDS-PAGE (CBB staining) (Fig.1)

Reaction Conditions
Incubate in 1 x Dnmt1 Reaction Buffer (20mM Tris-HCl pH7.4 0.5 mM EDTA 0.2 mM DTT 5% glycerol) with 10μM S-adenosylmethionine (SAM) at 37℃
 
 
Code
Quantity
                  Price
 
10-201
300 U
330 USD

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