Product Class: Other

Fpg
NEB1 recombinant Gray property icon for incubation temperature 37° 60 Heat Gray property icon for rAlbumin

Product Introduction

Diagram of mechanism

  • 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.

 

Bioz Badge Exists : True
Catalog # Size Concentration
M0240S 500 units 8000 units/ml
M0240L 2500 units 8000 units/ml

Product Information

Description

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)
This product is related to the following categories:
DNA Repair Enzymes and Structure-specific Endonucleases
This product can be used in the following applications:
Polymerases for DNA Manipulation

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 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: https://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

References

  1. Tchou, J. et al. (1994). Substrate specificity of Fpg protein. J. Biol.Chem.. 269, 15318-15324.
  2. Hatahet, Z. et al. (1994). Newsubstrates for old enzymes. J. Biol.Chem.. 269, 18814-18820.
  3. 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.
  4. 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.
  5. Collins, A., Duthie, S. and Dobson, V. (1993). Direct enzymatic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis. 14, 1733-1735.
  6. 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.
  7. 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.
  8. 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.
  9. 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.

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the activity of Fpg in the NEBuffers 1-4?
  2. Does Fpg cleave hydroxymethyl uracil?
  3. Does Fpg cut only one strand, or do they cause a double-strand break?
  4. What is the Molecular Weight of Fpg?
  5. What is the difference between using Fpg and hOOG1?
  6. Does the Fpg contain a tag?
  7. What buffer will work with both Endonuclease III and Fpg?
  8. What substrate is Fpg tested on?
  9. What is the activity of Fpg in rCutSmart Buffer?

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

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 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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