Product Class: Other

Uracil Glycosylase Inhibitor (UGI)
NEBU cloned at NEB recombinant 37 70 Heat

Product Introduction

  • Uracil Glycosylase Inhibitor is a small protein (9.5 kDa) which inhibits E. coli uracil-DNA glycosylase (UDG) as well as UDG from other species
  • Remains partially active following heat treatment at 95°C
  • Can be used to prevent subsequent degradation of product DNA
Catalog # Size Concentration
M0281S 200.0 units 2000 units/ml
M0281L 1000.0 units 2000 units/ml

Product Information

Description

The Uracil Glycosylase Inhibitor (UGI) of Bacillus subtilis bacteriophage PBS1 is a small protein (9.5 kDa) which inhibits E. coli uracil-DNA glycosylase (UDG) as well as UDG from other species. Inhibition of UDG occurs by reversible protein binding with a 1:1 UGD:UGI stoichiometry. UGI is capable of dissociating UDG-DNA complexes.

Product Source

An E. coli strain that carries the cloned UGI gene from Bacillus subtilis bacteriophage PBS1.
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
  • M0281S     -20    
      Uracil Glycosylase Inhibitor (UGI) M0281SVIAL -20 1 x 0.1 ml 2,000 units/ml
      UDG Reaction Buffer B0280SVIAL -20 1 x 1.5 ml 10 X
  • M0281L     -20    
      Uracil Glycosylase Inhibitor (UGI) M0281LVIAL -20 1 x 0.5 ml 2,000 units/ml
      UDG Reaction Buffer B0280SVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit of UGI is defined as the amount of protein required to inhibit one unit of E. coli UDG in 1 hour at 37°C in a total reaction volume of 50 µl. One unit of UDG is the amount of enzyme which will catalyze the release of 60 pmol of uracil per minute from double stranded, uracil-containing DNA.

Reaction Conditions

1X UDG Reaction Buffer
Incubate at 37°C

1X UDG Reaction Buffer
20 mM Tris-HCl
1 mM DTT
1 mM EDTA
(pH 8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

70°C for 10 minutes

Application Features

  • Uracil glycosylase inhibitor (UGI) inhibits uracil-DNA glycosylase (UDG). Since UDG remains partially active following heat treatment at 95°C, UGI can be used to prevent subsequent degradation of product DNA.

Product Notes

  1. Since UDG remains partially active following heat treatment at 95°C, UGI can be used to prevent subsequent degradation of product DNA.
  2. Works only on E. coli UDG (NEB #M0280).  
  3. Use equal or more units of UGI than units of UDG in the original reaction.
  4. Active in NEBuffers 1-4.  (NEB #B7001, NEB #B7002, NEB #B7003, NEB #B7004)
  5. UGI is extremely thermostable, retaining > 95% activity after boiling for 10 minutes.

References

  1. Lindahl, T. et al. (1977). J. Biol.Chem.. 252, 3286-3294.
  2. Wang, Z., et al. (1991). Gene. 99, 31-37.
  3. Bennett, S. E. and Mosbaugh, D. W. (1992). J. Biol. Chem.. 267, 22512-22521.

Protocols, Manuals & Usage

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. What is the nature of the Uracil Glycosylase Inhibitor?
  2. How does UGI inhibit UDG?
  3. Does UGI inhibit the activity of all UDGs?
  4. What is the molecular weight of UGI?
  5. Why is it necessary to use UGI in a reaction?

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.

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