Product Class: Other

Afu Uracil-DNA Glycosylase (UDG)
Gray property icon for NEBU Gray property icon for cloned at NEB recombinant 65 Gray property icon for heat inactivation no

Afu UDG has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10225518. Learn more.

Product Introduction

Diagram of mechanism

  • Archaeoglobus fulgidus Uracil DNA Glyocsylase
  • Thermostable monofunctional DNA glycosylase that catalyzes the hydrolysis of the N-glycosidic bond from deoxyuridine to release uracil
  • Active on ss and dsDNA
  • Heat inactivation not possible
Bioz Badge Exists : True
Catalog # Size Concentration
M0279S 200 units 2000 units/ml

Product Information

Description

A thermostable homolog of the E. coli Uracil-DNA Glycosylase (UDG) (1,2) from Archaeoglobus fulgidus. Afu UDG catalyzes the release of free uracil from uracil-containing DNA. Afu UDG efficiently hydrolyzes uracil from single-stranded or double-stranded DNA.

Product Source

Cloned and overexpressed in E. coli.
This product is related to the following categories:
DNA Repair Enzymes and Structure-specific Endonucleases,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    M0279S -20
    Component Name Component # Stored at (°C) Amount Concentration
    Afu Uracil-DNA Glycosylase (UDG) M0279SVIAL -20 1 x 0.1 ml 2,000 units/ml
    ThermoPol® II (Mg-free) Reaction Buffer B9005SVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to release 60 pmol per minute of a fluorescently labeled 47 mer single-stranded DNA oligonucleotide containing a single uracil base in 30 minutes at 65°C in a total reaction volume of 50 µl in 1X Thermopol II Buffer.

Reaction Conditions

1X ThermoPol® II (Mg-free) Reaction Buffer
Incubate at 65°C

1X ThermoPol® II (Mg-free) Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
0.1 mg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Molecular Weight

Theoretical: 22717 daltons

Specific Activity

96 units/mg

Unit Assay Conditions

1X ThermoPol II (Mg-free) Reaction Buffer, 1 unit of Afu Uracil-DNA Glycosylase, 0.2 µg 3H-uracil DNA (104-105 cpm/µg) for 30 minutes at 65°C in a total reaction volume of 50 µl.

Product Notes

  1. Afu UDG retains 50% activity in the presence of 150 mM NaCl. Afu UDG retains less than 1% activity after boiling for 30 minutes in standard reaction conditions.
  2. Under standard reaction conditions, uracil glycosylase inhibitor (UGI) does not inhibit Afu UDG.

References

  1. Sandigursky, M. and Franklin, W. A. (1999). CurrentBiology. 9,
  2. Sandigursky, M. and Franklin, W. A. (2000). J. Biol.Chem.. 275,

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Will Afu UDG work in Thermopol Buffer?
  2. What is the molecular weight of Afu UDG?
  3. Is Afu UDG a tagged protein?
  4. I see that Afu UDG works optimally at 65°C. Do you have another glycosylase that works at lower temperatures?
  5. Does the Uracil Glycosylase inhibitor (UGI) inhibit Afu UDG?
  6. Does Afu UDG release uracil from ss and dsDNA?
  7. Does Afu UDG cut RNA?
  8. Are there any specific recommendations for the use of UDG on single stranded DNA?
  9. Will Afu UDG work in Fpg buffer?
  10. What is the activity of WarmStart Afu Uracil-DNA Glycosylase (UDG) in NEBuffers?

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.

Specification Change Notifications

Afu UDG has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10225518. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

Effective 01/18/2024, unit definition has changed.

 

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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