Product Class: Other

Human DNA (cytosine-5) Methyltransferase (Dnmt1)
NEBU cloned at NEB recombinant 37 65 Heat BSA SAM Epi

  • Catalog # M0230 was discontinued on December 15, 2021
M0230cutsite_thumb

Product Introduction

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).

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Catalog # Size Concentration

Product Information

Description

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).
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0230S     -20    
      Human DNA (cytosine-5) Methyltransferase (Dnmt1) M0230SVIAL -20 1 x 0.025 ml 2,000 units/ml
      Dnmt1 Reaction Buffer B0230SVIAL -20 1 x 1.5 ml 10 X
      Purified BSA B9001SVIAL -20 1 x 0.25 ml 10 mg/ml
      S-adenosylmethionine (SAM) B9003SVIAL -20 1 x 0.1 ml 32 mM
  • M0230L     -20    
      Human DNA (cytosine-5) Methyltransferase (Dnmt1) M0230LVIAL -20 1 x 0.125 ml 2,000 units/ml
      Dnmt1 Reaction Buffer B0230SVIAL -20 1 x 1.5 ml 10 X
      Purified BSA B9001SVIAL -20 1 x 0.25 ml 10 mg/ml
      S-adenosylmethionine (SAM) B9003SVIAL -20 1 x 0.1 ml 32 mM

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

Product 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 & Troubleshooting

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?

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

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

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.