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Fpg

Catalog # Concentration Size List Price Quantity Your Price
M0240L 8000 units/ml 2500 units $494.00
$444.60
M0240S 8000 units/ml 500 units $122.00
$109.80
Catalog # Size List Price Your Price
M0240L 2500 units $494.00
$444.60
M0240S 500 units $122.00
$109.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • Formamidopyrimidine DNA Glycosylase
  • Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities
  • N-glycosylase activity releases damaged purines, including 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyG) and 8-oxo-7,8-dihydroguanine (8-oxoG), generating an AP site. The AP lyase activity cleaves an AP site, via β and δ-elimination, creating a 1 nucleotide DNA gap with 5' and 3' phosphate termini.

 

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    What are endonucleases and their applications?
Fpg (formamidopyrimidine [fapy]-DNA glycosylase) (also known as 8-oxoguanine DNA glycosylase) acts both as a N-glycosylase and an AP-lyase. The N-glycosylase activity releases damaged purines from double stranded DNA, generating an apurinic (AP site). The AP-lyase activity cleaves both 3´ and 5´ to the AP site thereby removing the AP site and leaving a 1 base gap. Some of the damaged bases recognized and removed by Fpg include 7, 8-dihydro-8-oxoguanine (8-oxoguanine), 8-oxoadenine, fapy-guanine, methy-fapy-guanine, fapy-adenine, aflatoxin B1-fapy-guanine, 5-hydroxy-cytosine and 5-hydroxy-uracil (1,2).
Product Source
An E.coli strain that carries the cloned fpg gene (3)
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Fpg M0240SVIAL -20 1 x 0.065 ml 8,000 units/ml
  NEBuffer™ 1 B7001SVIAL -20 1 x 1.25 ml 10 X
  Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
  Fpg M0240LVIAL -20 1 x 0.32 ml 8,000 units/ml
  NEBuffer™ 1 B7001SVIAL -20 1 x 1.25 ml 10 X
  Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 10 pmol of a 34-mer oligonucleotide duplex containing a single 8-oxoguanine base paired with a cytosine in a total reaction volume of 10 μl in 1 hour at 37°C.

Reaction Conditions

1X NEBuffer™ 1
Supplement with 100 µg/ml Recombinant Albumin, Molecular Biology Grade
Incubate at 37°C

1X NEBuffer™ 1
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
1 mM DTT
(pH 7 @ 25°C)

Dilution for Comet Assay

Recommended dilution for the Comet Assay: 1:103 to 1:104 (4,5,6). For a protocol please visit: http://cometassay.com.

Storage Buffer

20 mM Tris-HCl
50 mM NaCl
0.5 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 8 @ 25°C

Heat Inactivation

60°C for 10 minutes

Materials Sold Separately
References
  • Tchou, J. et al. (1994). Substrate specificity of Fpg protein. J. Biol.Chem.. 269, 15318-15324.
  • Hatahet, Z. et al. (1994). Newsubstrates for old enzymes. J. Biol.Chem.. 269, 18814-18820.
  • Boiteux, S., O'Connor, T. and Laval, J. (1987). Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein. EMBOJ.. 5, 18814-18820.
  • Singh, N., McCoy, M., Tice, R. and Schneider, L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Exp.Cell Res.. 175, 184-191.
  • Collins, A., Duthie, S. and Dobson, V. (1993). Direct enzymatic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis. 14, 1733-1735.
  • Collins, A., Dusinska, M., Gedik, C. and Stetina, R. (1996). Oxidative damage to DNA: do we have a reliable biomarker?. Environ.Health Perspect.. 104, 465-469.
  • Pflaum, M., Will, O., Mahler, H.-C. and Epe, B. (1998). DNA oxidation products determined with repair endonucleases in mammalian cells: types, basal levels and influence of cell proliferation. FreeRad. Res.. 29, 585-594.
  • Hartwig, A., Dally, H. and Schlepegrell, R. (1996). Sensitive analysis of oxidative DNA damage in mammalian cells: use of the bacterial Fpg protein in combination with alkaline unwinding. Toxicol.Lett.. 88, 85-90.
  • Czene, S. and Harms-Ringdahl, M. (1995). Detection of single strand breaks and formamidopyrimidine-DNA glycosylase-sensitive sites in DNA of cultured human fibroblasts. Mutat.Res.. 336, 235-242.
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.
Change Notifications
Effective April 24, 2023, Recombinant Albumin (rAlbumin) replaces BSA
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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