Product Class: Other

Thermostable OGG
Gray property icon for rCutSmart recombinant 65

Product Introduction

Mechanism of Thermostable OGG
  • Thermostable oxoguanine glycosylase
  • Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities
  • N-glycosylase activity releases 8-oxo-7,8-dihydroguanine (8-oxoG), generating an AP site. The AP-lyase activity cleaves 3′ to the AP site leaving a 5′ phosphate and a 3′-phospho-α, β-unsaturated aldehyde.
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Bioz Badge Exists : True
Catalog # Size Concentration
M0464S 500 units 8000 units/ml

Product Information

Description

Highlights

  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
Thermostable OGG is an archaeal 8-oxoguanine (8-oxoG) DNA glycosylase which acts both as a N-glycosylase and an AP-lyase. The N-glycosylase activity releases the damaged purine (8-oxoguanine) from double stranded DNA, generating an apurinic (AP) site. The AP-lyase activity cleaves 3′ to the AP site leaving a 5′ phosphate and a 3′-phospho-α, β-unsaturated aldehyde. Unlike some other DNA glycosylases, Thermostable OGG specifically recognizes and cleaves only 8-oxoG and no other modified bases.  

Figure 1: Unlike Fpg, Thermostable OGG specifically recognizes and cleaves only 8-oxoguanine and no other modified bases 

RFU of Thermostable OGG as compared to Fpg, showing cleaved product and substrate

The ability to excise modified bases was queried by incubating Thermostable OGG (8 U) or Fpg (8 U) with fluorescently labeled dsDNA containing modified bases (either 8-oxoadenine, 5-hydroxycytosine, thymine glycol, 5-hydroxyuridine or 8-oxoguanine). Samples were analyzed by capillary electrophoresis and cleavage products are evidenced by the appearance of a cleaved product peak. Thermostable OGG only cleaves the 8-oxoguanine sample (top panel) while Fpg is active on other substrates including , 5-hydroxycytosine, thymine glycol and  5-hydroxyuridine (bottom panel).

Product Source

An E. coli strain that carries a cloned thermostable archael oxoguanine glycosylase gene.
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

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 10 pmol of a 60-mer fluorescently labeled oligonucleotide duplex containing a single 8-oxoguanine base paired with a cytosine in a total reaction volume of 10 μl in 1 hour at 65°C.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 65°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
500 mM NaCl
0.1 mM EDTA
1 mM DTT
200 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the difference between Thermostable OGG (NEB #M0464) and Fpg (NEB #M0240)?
  2. What is the activity of Thermostable OGG in other buffers?
  3. Will Thermostable OGG cut an 8-oxoguanine paired with a base other than cytosine?
  4. What is the molecular weight of Thermostable OGG?
  5. Is Thermostable OGG a tagged protein?
  6. What substrate is used to test Thermostable OGG?
  7. Does Thermostable OGG cut only one strand, or both, to cause a double-strand break?
  8. What modified bases will Thermostable OGG recognize and cleave?
  9. Will Thermostable OGG cut 8-oxoguanine in ssDNA?
  10. What temperatures is Thermostable OGG active?

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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