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hAAG

Catalog # Concentration Size List Price Quantity Your Price
M0313S 10000 units/ml 500 units $123.00
$110.70
Catalog # Size List Price Your Price
M0313S 500 units $123.00
$110.70
Catalog #
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • 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)
Featured Videos
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  • Monarch_MiniprepProtocol_VideoThumb
    What are endonucleases and their applications?
Product Source
An E. coli strain which carries the cloned truncated human AAG gene (1)
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  hAAG M0313SVIAL -20 1 x 0.05 ml 10,000 units/ml
  ThermoPol® Reaction Buffer B9004SVIAL -20 1 x 1.5 ml 10 X
Application Features
  • Single cell gel electrophoresis (Comet Assay)(5,6,7) 
  • Alkaline elution (8)
  • Alkaline unwinding (9)

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.

Materials Sold Separately
Notes
  • hAAG has less than 25% activity in NEBuffers 1-4 (NEB #B7001, NEB #B7002, NEB #B7003, NEB #B7004)
References
  • O'Brien, P. and Ellenberger, T. (2003). Biochemistry. 42, 12418-12429.
  • Abner, C.W.et al. (2001). J. Biol. Chem.. 276, 13379-13387.
  • Samson, L. (1991). Proc. Natl. Acad.. Sci. 88, 9127-9131.
  • Lau, A.Y. et al. (1998). Cell. 95, 249-258.
  • Singh, N. et al. (1961). Experimental Cell  Research. 175, 184-191.
  • Collins, A. et al. (1993). Carcinogenesis . 14, 1733-1735.
  • Collins, A. et al. (1996). Environmental Health Perspectives. 104, 465-469.
  • Pflaum, M. et al. (1996). Free Rad. Res. 29, 585-594.
  • Hartwig, A. et al. (1996). Toxicology Letters.. 88, 85-90.
Protocols
  1. Protocol for hAAG (M0313)
Usage Guidelines
Quality Control Assay
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 Data Sheets
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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.

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