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Human DNA (cytosine-5) Methyltransferase (Dnmt1)

Catalog # Concentration Size List Price Quantity Your Price
M0230L 2000 units/ml 250 units
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M0230S 2000 units/ml 50 units
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Catalog # Size List Price Your Price
M0230L 250 units
M0230S 50 units
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Dnmt1 methylates cytosine residues in hemimethylated DNA in the sequence of 5'...CG...3' (1,2). Mammalian Dnmt1 is believed to be involved in carcinogenesis, embryonic development and several other biological functions (3-5).

The bulk of the methylation takes place during DNA replication in the S-phase of the cell cycle (6).
Product Source
Dnmt1 is expressed from human Dnmt1 cDNA using a baculovirus expression system (1,7).

Properties & Usage

Unit Definition

One unit is the amount of enzyme required to catalyze the transfer of 1 pmol of methyl group to poly dI.dC substrate in a total reaction volume of 25 μl in 30 minutes at 37°C.

Reaction Conditions

1X Dnmt1 Reaction Buffer
Supplement with 100 µg/ml BSA, Molecular Biology Grade and 160 µM S-adenosylmethionine (SAM)
Incubate at 37°C

Usage Concentration

2,000 units/ml

Storage Buffer

50 mM Tris-HCl
200 mM NaCl
1 mM DTT
1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

65°C for 20 minutes

Companion Products
Materials Sold Separately
Notes
  • For DNA modification and protection applications, M.SssI (NEB #M0226) is preferred because it efficiently methylates both unmethylated and hemimethylated DNA substrates.
References
  • Pradhan, S. et al. (1999). J. Biol. Chem. . 274, 33002-33010.
  • Bacolla, A. et al. (1999). J. Biol. Chem. . 274, 33011-33019.
  • Schmutte, C. et al. (1998). Biol. Chem. . 379, 377-388.
  • Laird, P.W. et al. (1995). Cell. 81, 197-205.
  • Li, E. et al. (1992). Cell. 12, 915-926.
  • Leonhardt, H. et al. (1992). Cell. 71, 865-873.
  • Yen, R.W. et al. (1992). Nucl. Acids. Res. . 20, 2287-2291.
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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