Human DNA (cytosine-5) Methyltransferase (Dnmt1)

Description

Dnmt1 methylates cytosine residues in hemimethylated DNA at 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).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
Dnmt1 Reaction Buffer10X
Purified BSA-2010 mg/ml
S-adenosylmethionine (SAM)-2032 mM

Properties and 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
Incubate at 37°C

Usage Concentration

2,000 units/ml

Storage Temperature

-20°C

Storage Conditions

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 min

Notes

  1. For DNA modification and protection applications, M.SssI (NEB #M0226) is preferred because it efficiently methylates both unmethylated and hemimethylated DNA substrates.

References

  1. Pradhan, S. et al. (1999). J. Biol. Chem. . 274, 33002-33010.
  2. Bacolla, A. et al. (1999). J. Biol. Chem. . 274, 33011-33019.
  3. Schmutte, C. et al. (1998). Biol. Chem. . 379, 377-388.
  4. Laird, P.W. et al. (1995). Cell. 81, 197-205.
  5. Li, E. et al. (1992). Cell. 12, 915-926.
  6. Leonhardt, H. et al. (1992). Cell. 71, 865-873.
  7. Yen, R.W. et al. (1992). Nucl. Acids. Res. . 20, 2287-2291.

FAQs

  1. Is S-adenosylmethionine (SAM) supplied with the Methyltransferase?
  2. What source of tritiated SAM is recommended for use with Dnmt1?
  3. What should be considered if the methylation using Dnmt1 is not going to completion?
  4. How does Dnmt1 differ from SssI methylase?
  5. Can Dnmt1 be heat inactivated?
  6. Will any NEB methyltransferases (methylases) work on RNA?

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Interactive Tools

Quality Control

Quality Assurance Statement

  • Purified free of contaminating endonucleases and exonucleases.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.