Product Class: Other

hAAG
NEBU cloned at NEB recombinant 37 65 Heat

The large size of this product has been discontinued.

Product Introduction

  • Human Alkyl Adenine DNA Glycosylase
  • Monofunctional DNA glycosylase that catalyzes the hydrolysis of the N-glycosidic bond to release damaged base
  • Recognizes a wide variety of alkylated and oxidized purines
  • Major substrates include 3-methyladenine, 7-methylguanine, 1,N6-ethenoadenine, and hypoxanthine
  • Minor substrates include deoxyinosine and deoxyxanthosine
  • Also known as methylpurine DNA glycosylase (MPG) and alkyl-N-purine DNA glycosylase (ANPG)
Catalog # Size Concentration
M0313S 500.0 units 10000 units/ml

Product Information

Description

Product Source

An E. coli strain which carries the cloned truncated human AAG gene (1)
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

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to create an AP site from 1 pmol of a 34-mer oligonucleotide duplex containing a single deoxyinosine site in a total reaction volume of 10 µl in 1 hour at 37°C. 

Reaction Conditions

1X ThermoPol® Reaction Buffer
Incubate at 37°C

1X ThermoPol® Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.5% Tween® 20
0.5% IGEPAL® CA-630
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Molecular Weight

Theoretical: 25752 daltons

Unit Assay Conditions

1X ThermoPol Buffer containing 5 pmol of fluorescently labeled oligonucleotide duplex in a total reaction volume of 10 µl.

Application Features

  • Single cell gel electrophoresis (Comet Assay)(5,6,7) 
  • Alkaline elution (8)
  • Alkaline unwinding (9)

Product Notes

  1. hAAG has less than 25% activity in NEBuffers 1-4 (NEB #B7001, NEB #B7002, NEB #B7003, NEB #B7004)

References

  1. O'Brien, P. and Ellenberger, T. (2003). Biochemistry. 42, 12418-12429.
  2. Abner, C.W.et al. (2001). J. Biol. Chem.. 276, 13379-13387.
  3. Samson, L. (1991). Proc. Natl. Acad.. Sci. 88, 9127-9131.
  4. Lau, A.Y. et al. (1998). Cell. 95, 249-258.
  5. Singh, N. et al. (1961). Experimental Cell  Research. 175, 184-191.
  6. Collins, A. et al. (1993). Carcinogenesis . 14, 1733-1735.
  7. Collins, A. et al. (1996). Environmental Health Perspectives. 104, 465-469.
  8. Pflaum, M. et al. (1996). Free Rad. Res. 29, 585-594.
  9. Hartwig, A. et al. (1996). Toxicology Letters.. 88, 85-90.

Protocols, Manuals & Usage

Protocols

  1. Protocol for hAAG (M0313)

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What substrate is used to test hAAG?
  2. What is the molecular weight of hAAG?
  3. Is hAAG a tagged protein?

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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